{"title":"Other Kit","description":null,"products":[{"product_id":"rnase-inhibitor-elisa-kit","title":"RNase Inhibitor ELISA Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e1. Preparation:\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e1. Before using all components of reagent and sample under test needs to be restored at room temperature.\u003cbr\u003e\u003cbr\u003e1 x for Wash Buffer preparation: 20 x will Wash Buffer balance to room temperature to crystallization completely dissolved, using deionized water or ultrapure water diluted 20 times after blending, diluted 1 x for Wash Buffer every hole are use 3 ml, in 2-8 ℃ can keep stable for 1 week.\u003cbr\u003e\u003cbr\u003e2. RNase Inhibitor Detection Antibody Dilution: before using blender, and then use Dilution Buffer 2 will RNase Inhibitor DetectionAntibody according to 1: 100 dilution ratio of dilution.\u003cbr\u003e\u003cbr\u003e3. Preparation of the substrate solution: Color Reagent A and Color Reagent B were mixed according to 1:1, and then mixed for immediate use. The dosage of each hole 100 mu L. Do not use if the mixed substrate solution turns blue.\u003cbr\u003e\u003cbr\u003e4. RNase Inhibitor dilution of Standard: 6μL RNase Inhibitor Standard stock solution (10μg\/mL) was diluted 100-fold with 594 μl dilution Buffer 1 as the first gradient (100ng\/mL). Then take 100 ng\/mL concentration sample 300 mu L to new EP tube, add 300 mu L Dilution Buffer1, double gradient Dilution, Dilution into 50, 25, 12.5, 6.25, 3.125 and 1.5625 ng\/mL, a total of seven levels.\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240312\/fce65833798446c3ac1092bef4cc23d0.png\" alt=\"\" width=\"603\" height=\"185\"\u003e\u003cbr\u003e\u003cbr\u003eIn order to ensure the validity of the experimental results, please use the newly prepared standard solution for each experiment.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e2, the operation process:\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e1. All operating at room temperature, it is recommended that the standard and the sample under test to determine double complex hole.\u003cbr\u003e\u003cbr\u003e1. sample incubation: will be diluted RNase Inhibitor of Standard and sample under test in enzyme label plate, respectively, 100 mu L\/hole, blending, cover the seal plate membrane, 37 ℃ for 2 h incubation.\u003cbr\u003e\u003cbr\u003e2. washing board: abandon to liquid in the hole, in 1 x for Wash Wash Buffer board, 300 mu L\/hole, soak 30 s, washing board 5 times, the last time to pat dry residual liquid.\u003cbr\u003e\u003cbr\u003e3. Inhibitor Detection Antibody incubation: after will dilute the RNase Inhibitor Detection Antibody in enzyme label plate, 100 mu L\/hole, blending, cover the seal plate membrane, 37 ℃ for 1 h incubation.\u003cbr\u003e\u003cbr\u003e4. Wash the plate: discard the liquid in the hole, wash the plate with 1×Wash Buffer, 300μL\/ well, soak for 30s, wash the plate 5 times, and pat the residual liquid dry as much as possible for the last time.\u003cbr\u003e\u003cbr\u003e5. Color development: The prepared substrate solution was added to the microplate, 100μL\/ well, mixed, covered with plate sealing membrane, and incubated at 37 ° C in the dark for 20min.\u003cbr\u003e\u003cbr\u003e6. Termination: Add Stop Reagent to the microplate, 50μL\/ well, and gently shake the microplate until the color is uniform to ensure that the color development time of each well is the same.\u003cbr\u003e\u003cbr\u003e7. reading value: enzyme label plate to be included in the enzyme standard instrument, read in 450 nm wavelength absorbance, read values within 20 minutes.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e3. Data processing\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003eIf the OD value of the sample to be tested exceeded the OD value of the highest point of the standard curve, the sample should be diluted and measured again.\u003cbr\u003e\u003cbr\u003eWith standard concentration as the abscissa, standard values for the vertical suction light drawing standard curve. ELISACalc.exe regression and fitting calculation software is recommended for four-parameter fitting to calculate the concentration of RNase Inhibitor in samples.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe kit is a double-antibody sandwich ELISA method, which is precoated with RNase Inhibitor to capture antibody. After adding the sample, the sample is captured to form antibody-antigen complex, and then HRP-labeled RNase Inhibitor is added to detect antibody. An antibody-antigen-antibody \"sandwich\" complex was formed. Finally, TMB was added for color development, and the absorbance (OD) value was read at 450nm wavelength after the reaction was terminated. The RNase Inhibitor content in the sample was positively correlated with the OD value.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eRNase Inhibitor can specifically inhibit the activities of RNase A, B and C, and can form a 1:1 complex with RNase, thereby inhibiting its activity. \u003cbr\u003e        In the process of drug mRNA vaccine production, RNase Inhibitor can effectively inhibit the reaction system of possible RNase activity, so as to protect the mRNA degradation. According to the regulation, need to concentrate to the residue detection of mRNA vaccine, including RNase Inhibitor as protein product is belong to the category of protein residue test items, so we need the residues were quantitative detection. \u003cbr\u003e        This kit, double antibody sandwich enzyme-linked immunoassay (ELISA) to detect RNase Inhibitor, with high sensitivity and specificity of detection and quantitative analysis of RNase Inhibitor content.\u003cbr\u003e\u003cbr\u003eProduct Features: \u003cbr\u003eSensitivity: 3.125 ng\/ml \u003cbr\u003eDetection Range: 3.125 100 ng\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003ctable style=\"border-collapse: collapse; width: 100%; height: 154px;\" border=\"1\" cellspacing=\"1\" cellpadding=\"0\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; width: 188pt; text-align: center;\" width=\"376\" height=\"29\"\u003eComponents\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 154pt; height: 14px; text-align: center;\" width=\"308\"\u003eDescriptions\u003c\/td\u003e\n\u003ctd class=\"et3\" style=\"width: 68.5pt; height: 14px; text-align: center;\" width=\"137\"\u003eSize\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eRNase Inhibitor Microplate\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003ePackage is capture antibody\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e96T\/Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eRNase Inhibitor Detection Antibody\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003eHRP was labeled and the dilution ratio was 1:100\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e150μl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eRNase Inhibitor Standard\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003eRestructuring RNase inhibitor (5 mu g\/ml)\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e150μl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003e20×Wash Buffer\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003escrubbing solution\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e30ml×2\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eSample Dilution Buffer\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003eSample diluent\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e30ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eConjugate Dilution Buffer\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003eenzyme diluent\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e15ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eColor Reagent A\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003eColor liquid A\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e6ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eColor Reagent B\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003eColor liquid B\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e6ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eStop Reagent\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003estop buffer\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e6ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.50pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\" height=\"29\"\u003eSealing Film\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003esealing film\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center;\"\u003e4 \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1, before the experiment should be in accordance with the requirements of relevant experimental reagent balance to room temperature. \u003cbr\u003e2, each reagent should be fully mixed before use to ensure the uniformity and accuracy of the reagent. \u003cbr\u003e3, strictly control the reaction temperature and reaction time. \u003cbr\u003e4, measurements can not beyond the normal range. \u003cbr\u003e5, the number of washing plate, the amount of washing plate liquid, the soaking time of washing plate liquid, and the strength of washing plate are the problems that should be paid attention to. \u003cbr\u003e6, when the termination reaction to avoid microporous of bubbles. \u003cbr\u003e7. The corresponding standard curve should be prepared for each test, and the standard curve of different kits and different days should not be mixed. \u003cbr\u003e8, pay attention to the change in time between different samples and steps with tray and spear, avoid cross contamination. \u003cbr\u003e9, reading the light absorption value should be completed within 20min after adding the termination solution. \u003cbr\u003e10, color with high concentration may produce black floc, belongs to the normal phenomenon, lightly don't affect the reading result.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eDry and keep away from light at 2-8 ° C, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e3.125-100 ng\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293314687051,"sku":"abs520031-96T","price":1183.0,"currency_code":"USD","in_stock":true}]},{"product_id":"cho-hcp-elisa-kit","title":"CHO HCP ELISA Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e1. Reagent preparation:\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e1. PBST 0.05% 15ml 20 × PBST 0.05%, dilute in ddH2O, dilute to 300ml.\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e2. 1.0% BSA Dissolve 1g of BSA in the bottle in 100ml PBST 0.05%, mix thoroughly until completely dissolved, and store at 2-8 °C. The prepared dilution buffer is valid for 7 days. It is recommended to prepare it as needed.\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e3. Test solution 2μg\/mL Take 48 μL of 0.5 mg\/mL Anti-CHO HCP-HRP and dilute it in 11952 μL of 1% BSA to obtain a final concentration of 2μg\/mL test solution.\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e4. Preparation of QC and CHOK1 HCP standards\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 97.8208%; height: 264px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 44px;\"\u003e\n\u003ctd style=\"width: 6.61224%; height: 44px; text-align: center;\"\u003eTube number\u003c\/td\u003e\n\u003ctd style=\"width: 9.85568%; height: 44px; text-align: center;\"\u003eDrug substance\u003c\/td\u003e\n\u003ctd style=\"width: 14.4932%; height: 44px; text-align: center;\"\u003eConcentration (ng\/mL)\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; height: 44px; text-align: center;\"\u003eVolume (μL)\u003c\/td\u003e\n\u003ctd style=\"width: 17.903%; height: 44px; text-align: center;\"\u003eVolume of 1% BSA (μL)\u003c\/td\u003e\n\u003ctd style=\"width: 10.1569%; height: 44px; text-align: center;\"\u003eTotal Volume\u003c\/td\u003e\n\u003ctd style=\"width: 15.7027%; height: 44px; text-align: center;\"\u003eFinal Concentration (ng\/mL)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 6.61224%; height: 22px; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 9.85568%; height: 22px; text-align: center;\"\u003eStandard\u003c\/td\u003e\n\u003ctd style=\"width: 14.4932%; height: 22px; text-align: center;\"\u003e0.2 mg\/mL\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; height: 22px; text-align: center;\"\u003e10\u003c\/td\u003e\n\u003ctd style=\"width: 17.903%; height: 22px; text-align: center;\"\u003e490\u003c\/td\u003e\n\u003ctd style=\"width: 10.1569%; height: 22px; text-align: center;\"\u003e500\u003c\/td\u003e\n\u003ctd style=\"width: 15.7027%; height: 22px; text-align: center;\"\u003e10000\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 6.61224%; height: 22px; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 9.85568%; height: 22px; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 14.4932%; height: 22px; text-align: center;\"\u003e10000\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; 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height: 22px; text-align: center;\"\u003e75\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; height: 22px; text-align: center;\"\u003e200\u003c\/td\u003e\n\u003ctd style=\"width: 17.903%; height: 22px; text-align: center;\"\u003e175\u003c\/td\u003e\n\u003ctd style=\"width: 10.1569%; height: 22px; text-align: center;\"\u003e375\u003c\/td\u003e\n\u003ctd style=\"width: 15.7027%; height: 22px; text-align: center;\"\u003e40\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 极 6.61224%; height: 22px; text-align: center;\"\u003eS4\u003c\/td\u003e\n\u003ctd style=\"width: 9.85568%; height: 22px; text-align: center;\"\u003eS3\u003c\/td\u003e\n\u003ctd style=\"width: 14.4932%; height: 22px; text-align: center;\"\u003e40\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; height: 22px; text-align: center;\"\u003e150\u003c\/td\u003e\n\u003ctd style=\"width: 17.903%; height: 22px; text-align: center;\"\u003e350\u003c\/td\u003e\n\u003ctd style=\"width: 10.1569%; height: 22px; text-align: center;\"\u003e500\u003c\/td\u003e\n\u003ctd style=\"width: 15.7027%; height: 22px; text-align: center;\"\u003e12\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 6.61224%; height: 22px; text-align: center;\"\u003eS5\u003c\/td\u003e\n\u003ctd style=\"width: 9.85568%; height: 22px; text-align: center;\"\u003eS4\u003c\/td\u003e\n\u003ctd style=\"width: 14.4932%; height: 22px; text-align: center;\"\u003e12\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; height: 22px; text-align: center;\"\u003e200\u003c\/td\u003e\n\u003ctd style=\"width: 17.903%; height: 22px; text-align: center;\"\u003e200\u003c\/td\u003e\n\u003ctd style=\"width: 10.1569%; height: 22px; text-align: center;\"\u003e400\u003c\/td\u003e\n\u003ctd style=\"width: 15.7027%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 6.61224%; height: 22px; text-align: center;\"\u003eS6\u003c\/td\u003e\n\u003ctd style=\"width: 9.85568%; height: 22px; text-align: center;\"\u003eS5\u003c\/td\u003e\n\u003ctd style=\"width: 14.4932%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; height: 22px; text-align: center;\"\u003e200\u003c\/td\u003e\n\u003ctd style=\"width: 17.903%; height: 22px; text-align: center;\"\u003e200\u003c\/td\u003e\u0026lt; td style = \"width: 10.1569%; height: 22 px; text-align: center; \"\u0026gt;400\u003ctd style=\"width: 15.7027%; height: 22px; text-align: center;\"\u003e3\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 6.61224%; height: 22px; text-align: center;\"\u003eNC\u003c\/td\u003e\n\u003ctd style=\"width: 极 9.85568%; height: 22px; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 14.4932%; height: 22px; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; height: 22px; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 17.903%; height: 22px; text-align: center;\"\u003e200\u003c\/td\u003e\n\u003ctd style=\"width: 10.1569%; height: 22px; text-align: center;\"\u003e200\u003c\/td\u003e\n\u003ctd style=\"width: 15.7027%; height: 22px; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 6.61224%; height: 22px; text-align: center;\"\u003eQC\u003c\/td\u003e\n\u003ctd style=\"width: 9.85568%; height: 22px; text-align: center;\"\u003eS1\u003c\/td\u003e\n\u003ctd style=\"width: 14.4932%; height: 22px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 10.7634%; height: 22px; text-align: center;\"\u003e50\u003c\/td\u003e\n\u003ctd style=\"width: 17.903%; height: 22px; text-align: center;\"\u003e200\u003c\/td\u003e\n\u003ctd style=\"width: 10.1569%; height: 22px; text-align: center;\"\u003e250\u003c\/td\u003e\n\u003ctd style=\"width: 15.7027%; height: 22px; text-align: center;\"\u003e20\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cstrong\u003e2. Required instruments:\u003c\/strong\u003e\u003cbr\u003e\u003ctable style=\"width: 65.3753%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center; width: 20.7692%;\"\u003e\u003cp\u003eConsumables\/Equipment\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center; width: 20.6793%;\"\u003emanufacturer\u003c\/td\u003e\n\u003ctd style=\"text-align: center; width: 49.1834%;\"\u003emodel\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center; width: 20.7692%;\"\u003e\u003cp\u003eMicroplate reader\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center; width: 20.6793%;\"\u003e\n\u003cp\u003eMolecular\u003c\/p\u003e\n\u003cp\u003eDevices\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; width: 49.1834%;\"\u003e\u003cp\u003eSpectra Max M5, M5e, Or equivalent\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center; width: 20.7692%;\"\u003e\u003cp\u003eThermostatic mixer\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center; width: 20.6793%;\"\u003e\u003cp\u003eEppendorf\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center; width: 49.1834%;\"\u003e\u003cp\u003eEppendorf\/5355, or equivalent\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center; width: 20.7692%;\"\u003e\u003cp\u003eVortex mixer\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center; width: 20.6793%;\"\u003e\u003cp\u003eIKA\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center; width: 49.1834%;\"\u003e\u003cp\u003eMS3 Digital, or equivalent\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e\u003cstrong\u003e3. Experimental process\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e1. Prepare the reagents according to the above \"reagent preparation\".\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e2. Take 50μL of standard, sample and QC on the well plate, then add 100μL of test solution (2μg\/mL), seal the labeled plate with a sealing membrane, place the labeled plate on the mixer, incubate at 500 rpm, 25 ± 3 ℃ for 2 hours.\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e3. Turn the microplate upside down in the water tank and discard the coating liquid. Take 300 μL of PBST 0.05% and wash the plate in microwells and discard the solution, and repeat the wash three times. Snap the board upside down on a clean paper towel and pat dry.\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e4. Add 100μL of TMB substrate to each well, seal the labeled plate, and incubate at 25 ± 3 ℃ in the dark for 15 minutes.\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e5. Add 100 μL of stop solution to each well.\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e6. Read the absorbance at the wavelength of 450\/650 nm with the microplate reader.\u003c\/strong\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e7. Analyze the data by SoftMax or similar software, and draw the standard curve with four-parameter fitting regression model.\u003c\/strong\u003e\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231114\/7682e47bdaec46e6b573ee44703443dc.png\" alt=\"\" width=\"392\" height=\"295\"\u003e\u003c\/p\u003e\n\u003cp\u003eNote: If the concentration of HCP in the sample exceeds the upper limit of the standard curve, it needs to be properly diluted with dilution buffer prior to testing.\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis method uses one-step enzyme-linked immunosorbent ELISA. Samples containing CHOK1 HCP can be simultaneously reacted with HRP-labeled goat anti-CHOK1 antibodies and anti-CHOK1 antibodies coated on enzyme plates to finally form a sandwich complex of solid phase antibodies-HCP-labeled antibodies. Unbound antigen antibodies can be removed by washing the plate. After the TMB substrate is added to the well to complete the reaction, the color development is stopped after the stop solution is added, and the OD or absorbance value of the reaction solution at 450\/650 nm is read with a microplate reader. The OD value or absorbance value is proportional to the HCP content in the solution. From this, the HCP concentration in the solution can be calculated from the standard curve.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eHost Cell Protein (HCP) secreted by CHO (Chinese Hamster Ovary) host cells is likely to cause contamination of biological products. Failure to adequately remove contaminants in the early stage of drug development may lead to a decrease in drug efficacy or induce an immune response. The kit can quantitatively detect the residue of host cell protein in bioengineering pharmaceuticals with high sensitivity, optimize the process flow and ensure the safety of biological products.\u003cbr\u003eProduct parameters:\u003cbr\u003e1. Sensitivity: 1ng\/mL\u003cbr\u003e2. Detection range: 3-100ng\/mL\u003cbr\u003e3. Precision: intra-batch coefficient of variation ≤ 10%, inter-batch coefficient of variation ≤ 15%\u003cbr\u003e4. HCP coverage: more than 80%\u003cbr\u003e5. Specificity: This kit is universal because it can specifically react with CHOK1 HCP without relying on the purification process\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003ctable style=\"width: 77.8137%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003eSerial number\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003eform\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003eConcentration\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003eStorage conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003eCHOK1 HCP Standard\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003e0.5 mg\/mL\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003e70uL\/tube\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003e≤-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003eAnti-CHO HCP-HRP\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003e0.5 mg\/mL\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003e60uL\/tube\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003e≤-20 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003eTMB\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003e12mL\/vial\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003e20 × PBST 0.05%\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003e15mL\/vial\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003e5\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003eStop liquid\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003e12mL\/vial\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003eEnzyme labeled plate sealing membrane\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003e3 sheets\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003e7\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003eBSA\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003e1g\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.6113%; text-align: center;\"\u003e8\u003c\/td\u003e\n\u003ctd style=\"width: 28.5372%; text-align: center;\"\u003eHigh adsorption 96-well prediction enzyme plate\u003c\/td\u003e\n\u003ctd style=\"width: 14.4398%; text-align: center;\"\u003eNA\u003c\/td\u003e\n\u003ctd style=\"width: 14.985%; text-align: center;\"\u003e1 piece\u003c\/td\u003e\n\u003ctd style=\"width: 17.286%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe termination solution is 2M sulfuric acid, please handle it carefully to prevent splashes!\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis kit is used to quantitatively detect the residual content of CHOK1 host cell protein in a sample.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eComponents 1 and 2 are stored at ≤-20 °C; Components 5 and 6 are stored at room temperature; Components 3, 4, 7, 8 are stored at 2-8 °C. Valid for 12 months.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293314785355,"sku":"abs590005-96T","price":952.0,"currency_code":"USD","in_stock":true}]},{"product_id":"kanamycin-residue-detection-kit","title":"Kanamycin Residue Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e I. Preparation: \u003c\/strong\u003e\u003cbr\u003e1 Kit preparation  \u003cbr\u003e Each component of the kit was allowed to equilibrate at room temperature  30min  After starting the operation.  \u003cbr\u003e2 Consumables and equipment that need to be provided by yourself  \u003cbr\u003e （ 1 ) microplate reader, thermostatic shaker (or thermostatic incubator), plate washer, vortex shaker, timer;  \u003cbr\u003e （ 2 ) High-precision pipettes and disposable tips ( 0.5-10uL 、 10-100uL 、 30-300uL 、 100-1000uL ）；  \u003cbr\u003e （ 3 ) deionized water (used to prepare the lotion ( 1× ））；  \u003cbr\u003e （ 4 ) absorbent paper, EP  tubes, disposable gloves;  \u003cbr\u003e3 Reagent preparation  \u003cbr\u003e （ 1 ) Lotion ( 1× ) Formulation  \u003cbr\u003e Take the lotion ( 20× ） 1  Portions, plus  19  Part of deionized water to prepare a working concentration washing solution ( 1× ）。 Lotion ( 20× If there is crystal formation, it should be placed at room temperature or  37℃ Shake gently in the water bath, and then dilute after the crystals are completely dissolved. Unused lotion ( 20× ) should be placed in  2-8℃ Storage.  \u003cbr\u003e （ 2 ) Preparation of Color Developing Solution  \u003cbr\u003e Chromogenic solution  A Chromogenic liquid  B  Mix in equal volumes, mix well and place in the dark. (Note: The time should not be left for too long, generally before use  10min  Formulation. Do not use if the mixed developer has turned blue).  \u003cbr\u003e （ 3 ) Preparation of standards  \u003cbr\u003e Dilute the standard with the sample diluent to  5ng\/mL And then use  2.5  The standard was prepared by dilution ratio (freshly prepared standard solution was used for each experiment) as shown below: \u003cdiv\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231023\/04cfc00131194e7c8a30f2dbb0a2d230.png\" alt=\"\" width=\"401\" height=\"181\"\u003e\u003c\/div\u003e4 Sample preparation  \u003cbr\u003e Restore the sample to room temperature and mix the sample before adding the sample; If the user needs to dilute the sample or the matching high-concentration standard in the box, the sample diluent in the box can be used for dilution; If it is a cell type sample, it is recommended to press  3000rpm\/min  Speed, centrifugal  5min Take the supernatant for detection. \u003cbr\u003e \u003cbr\u003e\u003cstrong\u003e 2. Operation process: \u003c\/strong\u003e\u003cbr\u003e1 Operation diagram \u003cdiv\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto; page-break-inside: avoid;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231023\/92b78c2016f645108c0a175b8e3fe49a.png\" alt=\"\" width=\"412\" height=\"510\"\u003e\u003c\/div\u003e2 Operation detailed steps  \u003cbr\u003e （ 1 ) Restore each component of the kit to room temperature  30min Take out the enzyme-labeled slats required for the test from the aluminum foil bag that has been equilibrated to room temperature, mark the order of the slats with a marker pen (it is recommended to perform double-well measurement), seal the remaining slats with a plate sealing film, put them back into the aluminum foil bag, seal them well, and place them in  2-8℃ Save. (Note: The slats are easy to fall off during the subsequent clapping process, so pay attention to marking them) \u003cbr\u003e （ 2 ) Sample incubation: Add to each well  50uL  Standard \/ Blank (sample dilution) \/ Sample, after which added  50uL  The antibody working solution was sealed with a plate sealing membrane and then placed in  25℃ Stationary incubation in the dark  30min 。 (Note: The standard must be added first. If the antibody is added first, it will react directly with the antigen on the plate; If the plate is not sealed or the plate is incompletely sealed during the incubation process, the reaction solution will evaporate, resulting in errors in the experiment; All incubation processes should be avoided as much as possible. Light irradiation)  \u003cbr\u003e （ 3 ) Wash the plate: After the incubation is completed, carefully remove the plate sealing membrane, discard the liquid in the well, and use the washing solution ( 1× ) Wash the plate  3  Second ( 250uL\/ well), and pat dry the residual liquid in the sample well. (Note: If hand washing board is used, add washing solution ( 1× ) When it is necessary to suspend liquid, it is best not to touch the inner wall of the hole; Each addition of the lotion ( 1× ) after resting  30s  (under the last slight shake of the standing process); When patting dry, be careful to change the absorbent paper each time or pat dry on a clean area of the paper)  \u003cbr\u003e （ 4 ) Enzyme-labeled secondary antibody incubation: enzyme-labeled secondary antibody is added to each well, 100uL\/ Well, seal the plate with a plate sealing film, and then place it in  25℃ Stationary incubation in the dark  30min 。  \u003cbr\u003e （ 5 ) Wash the plate: the method is the same as the procedure   （ 3 ）。  \u003cbr\u003e （ 6 ) Color development: press the pre-configured color developing liquid  100uL\/ The wells are added into the enzyme label plate, and the plate is sealed with a plate sealing membrane, 25℃ Stationary incubation in the dark  15min 。  \u003cbr\u003e （ 7 ) Termination: Adding a termination solution, 100uL\/ Holes, which can be read after uniform color development. (Note: It is recommended to set in the microplate reader reading program  5-10s  Oscillation of)  \u003cbr\u003e （ 8 ) Reading: Put the enzyme label plate into the microplate reader and set the wavelength to double wavelength  450\/630nm , read the absorbance value, the measurement should be after termination  20min  Inside completion. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e III. Data processing: \u003c\/strong\u003e\u003cbr\u003e1 Calculation of absorbance value  \u003cbr\u003e The absorbance value of each standard or sample is OD450nm-OD630nm 。  \u003cbr\u003e2 Calculation of percent absorbance  \u003cbr\u003e Mean absorbance value (double well) for each standard or sample divided by 0ng\/mL The mean absorbance value of the standard is multiplied by 100% That is, the percent absorbance:  \u003cbr\u003e Percentage absorbance ( % ）＝ B\/B0×100% \u003cbr\u003eB : Mean absorbance value of standard or sample  \u003cbr\u003eB0 ： 0ng\/mL Mean absorbance values of standards  \u003cbr\u003e3 Drawing and calculation of standard curve  \u003cbr\u003e Take the percentage absorbance value of the standard as the ordinate Y The standard concentration value is in the abscissa X , draw a standard curve, recommended using four parameters Logistic Mathematical model fitting equation:  \u003cbr\u003eY=((A-D)\/(1+(X\/C)^B))+D \u003cbr\u003e Substitute the percent absorbance value of the sample into the standard curve, read out the corresponding concentration value of the sample, and multiply it by its corresponding dilution factor to obtain the actual concentration of kanamycin in the sample. \u003cdiv\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231023\/556e8013101f475db19b3dbb7822775b.png\" alt=\"\" width=\"404\" height=\"220\"\u003e\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis kit uses an indirect competitive ELISA method to determine trace residues of kanamycin in samples. The enzyme label plate is pre-coated with conjugated antigen, and the residual kanamycin in the sample and the conjugated antigen pre-coated on the microwell strip compete for anti-kanamycin antibody. The enzyme-labeled secondary antibody is added, and then the color is developed by adding TMB substrate. The absorbance (OD value) is measured at the wavelength of 450 nm\/630 nm using a microplate reader, and the percent absorbance is calculated. The concentration of kanamycin in the sample is negatively correlated with the percent absorbance.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e         Kanamycin is an aminoglycoside antibiotic, which is used in the treatment of animal diseases, strain screening and preparation of drug raw materials for cell and gene therapy. 8 To the cranial nerves, resulting in vestibular and cochlear damage. The residues in animal food and biological drugs can affect human health and even cause allergic reactions. European and American countries and China all require its limited use. \u003cbr\u003e         Kanamycin quantitative assay kit uses indirect competition ELISA Methods, the pre-coated conjugated kanamycin antigen on the microwell strip competes with the residual kanamycin in the sample to bind to the enzyme-labeled anti-kanamycin monoclonal antibody, and then the enzyme-labeled secondary antibody is added TMB The substrate developed color, and the absorbance value was measured using a microplate reader, which was negatively correlated with the content of kanamycin in the sample. The operation time of this kit is only about one hour, and the linear range is 0.05ng\/mL-5ng\/mL 。 \u003cbr\u003e         When performing the linearity verification of kanamycin raw material, it is recommended that the kanamycin raw material be diluted to the linear range of the kit before determination, and diluted to at least a certain proportion  5  There was a linear correlation between the detection concentration and the dilution factor. \u003cbr\u003e         Specificity should be validated for other substances present in the process. It is recommended to use high concentration and low concentration quality control samples and add increasing concentrations of related interfering substances for specificity investigation. The matrix without the addition of analytes should also be measured simultaneously. The accuracy of the controls should be within a certain range and the measurements of the matrix without the addition of analytes should be below the lower limit of quantification. \u003cbr\u003e\u003cstrong\u003e Product Features: \u003c\/strong\u003e\u003cbr\u003e Strong specificity: No cross-reaction with penicillin, streptomycin sulfate, gentamicin and ampicin \u003cbr\u003e High Sensitivity: Kit sensitivity is 0.05ng\/mL\u003cbr\u003e Convenient and efficient: Using a two-step method, only 1 Results can be obtained in about hours \u003cbr\u003e Excellent performance: good standard curve linearity, high accuracy and good repeatability \u003cbr\u003e\u003cstrong\u003e Product performance indicators   ： \u003c\/strong\u003e\u003cbr\u003e Limit of detection: \u0026lt; 0.05 ng\/mL \u003cbr\u003e Limit of Quantification: 0.05 ng\/mL \u003cbr\u003e Linear Range: 0.05-5 ng\/mL \u003cbr\u003e Accuracy (spiked recovery): 70%-130% \u003cbr\u003e Accuracy (measurement deviation): ≤15% \u003cbr\u003e Repeatability (intra-batch difference): ≤15% \u003cbr\u003e\u003cstrong\u003e Specificity: \u003c\/strong\u003e\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 76.8647%; height: 86px; margin-left: auto; margin-right: auto;\" border=\"1\" width=\"60%\" cellspacing=\"0\" cellpadding=\"0\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; width: 38.9317%; text-align: center;\" width=\"221\" height=\"28\"\u003e\u003cstrong\u003e\u003cspan class=\"font1\"\u003e antibiotics \u003c\/span\u003e\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"width: 32.8242%; text-align: center; height: 14px;\" width=\"297\"\u003e\u003cstrong\u003e Cross reactivity rate \u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 16px; width: 38.9317%; text-align: center;\" height=\"28\"\u003e Kanamycin \u003c\/td\u003e\n\u003ctd class=\"et7\" style=\"width: 32.8242%; text-align: center; height: 16px;\"\u003e100%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et3\" style=\"height: 14px; width: 38.9317%; text-align: center;\" height=\"28\"\u003e Streptomycin sulfate \u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"width: 32.8242%; text-align: center; height: 14px;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et4\" style=\"height: 14px; width: 38.9317%; text-align: center;\" height=\"28\"\u003e gentamicin \u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"width: 32.8242%; text-align: center; height: 14px;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et3\" style=\"height: 14px; width: 38.9317%; text-align: center;\" height=\"28\"\u003e penicillin \u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"width: 32.8242%; text-align: center; height: 14px;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et5\" style=\"height: 14px; width: 38.9317%; text-align: center;\" height=\"28\"\u003e Ampicillin \u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"width: 32.8242%; text-align: center; height: 14px;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003ctable style=\"border-collapse: collapse; width: 94.7076%; height: 154px; border-color: #000000; border-style: solid; margin-left: auto; margin-right: auto;\" border=\"1\" width=\"95%\" cellspacing=\"0\" cellpadding=\"0\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; width: 12.0045%; text-align: center;\" width=\"96\" height=\"28\"\u003e\u003cstrong\u003e serial number \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 33.8945%; text-align: center; height: 14px;\" width=\"252\"\u003e\u003cstrong\u003e Name \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 21.2892%; text-align: center; height: 14px;\" width=\"139\"\u003e\u003cstrong\u003e Specifications \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.0942%; text-align: center; height: 14px;\" width=\"274\"\u003e\u003cstrong\u003e preservation \u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e1\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Enzyme label plate \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e8×12  Article \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃ Protect from light \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e2\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Standards ( 50 ng\/mL ） \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e1mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃ Protect from light \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e3\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Antibody working fluid \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e7mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e4\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Enzyme-labeled secondary antibody \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e12mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e5\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Sample dilution \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e30mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e6\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Lotion ( 20× ） \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e30mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e7\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Chromogenic solution  A\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e8mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃ Protect from light \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e8\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Chromogenic solution  B\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e8mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃ Protect from light \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e9\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Stop fluid \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e15mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 14.00pt;\"\u003e\n\u003ctd class=\"et2\" style=\"height: 14px; text-align: center; width: 12.0045%;\" height=\"28\"\u003e10\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 33.8945%; height: 14px;\"\u003e Plate sealing film \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 21.2892%; height: 14px;\"\u003e3 Zhang \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 20.0942%; height: 14px;\"\u003e-\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. The optimal reaction temperature of this kit is 25℃. If the temperature is too high or too low, the detection absorbance value and sensitivity will change.  \u003cbr\u003e2. All components in the kit must be restored to room temperature (20-25℃) before use.  \u003cbr\u003e3. Mix all components well before use, and the standard product needs to be briefly centrifuged for 5 seconds to concentrate all the liquid on the tube wall and cover at the bottom of the tube; Return all reagents to 2-8°C immediately after use.  \u003cbr\u003e4. The kit must be used within the shelf life, and the corresponding standard curve must be prepared for each test. It is not recommended to use different batches of relevant reagents in mixed batches.  \u003cbr\u003e5. Be careful not to touch the bottom of the microplate when adding liquid to the enzyme label plate to prevent damage to the coating layer. Replace the sampling tank and suction tip in time between different samples and steps to avoid cross-contamination.  \u003cbr\u003e6. When the slats are washed and dried, pay attention to prevent the slats from falling off, and the sealing film should not be reused.  \u003cbr\u003e7. High concentration may produce black flocs during color development, which is a normal phenomenon, and the slight degree does not affect the final reading result.  \u003cbr\u003e8. When reading, pay attention to whether the detection wavelength and fitting equation are selected correctly.  \u003cbr\u003e9. Only by strictly following the operation methods of the instructions and using all the reagents matched with this kit can the best detection effect be ensured.  \u003cbr\u003e10. The difference in test results can be caused by a variety of factors, including the operation of the experimenter, the use of the pipette, the washing technology, the reaction time or temperature, the storage of the kit, etc.  \u003cbr\u003e11. Our company is only responsible for the kit itself, not for the sample consumption caused by the use of the kit. Users are requested to fully consider the possible usage of samples before use and reserve sufficient samples.  \u003cbr\u003e12. The termination solution in this kit is an acid solution, so special attention should be paid to during operation.\u003cbr\u003e13. Operators should wear personal protective equipment, such as laboratory coats, gloves, masks and goggles.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eThe kit should be stored at 2-8℃, protected from light, with a shelf life of 12 months. Note that the unused kits after opening should still be stored at 2-8℃ in the dark.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293314916427,"sku":"abs590010-96T","price":663.0,"currency_code":"USD","in_stock":true}]},{"product_id":"gentamicin-residue-detection-kit","title":"庆大霉素残留检测试剂盒","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e 1. \u003c\/strong\u003e\u003cstrong\u003e Preparation  \u003cbr\u003e\u003c\/strong\u003e1 Kit preparation \u003cbr\u003e Each component of the kit was left to equilibrate at room temperature  30min  After starting the operation. \u003cbr\u003e2 、   Consumables and equipment required to be brought by yourself \u003cbr\u003e （ 1 ) Microplate reader, constant temperature oscillator (or constant temperature incubator), plate washing machine, vortex oscillator,   Timer; \u003cbr\u003e （ 2 ) High-precision pipettes and disposable tips ( 0.5-10µL 、 10-100µL 、 30-300µL 、  100-1000µL ）； \u003cbr\u003e （ 3 ) deionized water (used to prepare lotions ( 1× ））； \u003cbr\u003e （ 4 ) absorbent paper, EP  Tubes, disposable gloves. \u003cbr\u003e3 Reagent preparation \u003cbr\u003e （ 1 ) Lotion ( 1× ) formulation of \u003cbr\u003e Take the wash ( 20× ） 1  Portions, add  19  Parts of deionized water are prepared into a working concentration washing solution ( 1× ）。 Lotion ( 20× If crystals are formed in), it should be placed at room temperature or  37℃ Shake gently in the water bath and wait until the crystals are completely dissolved before diluting. Unused lotion ( 20× ) should be placed in  2-8℃ Storage. \u003cbr\u003e （ 2 ) Preparation of chromogenic solution \u003cbr\u003e The colored solution  A Chromogenic Solution  B  Mix in equal volumes, mix well and place in the dark. (Note: The time cannot be left for too long, before general use  10min  Formulation. Do not use if the mixed color solution has turned blue). \u003cbr\u003e （ 3 ) Preparation of standards \u003cbr\u003e Dilute the standard with sample diluent to  10ng\/mL , and then use  2.5  The standard is prepared by double dilution method (freshly prepared standard solution is used for each experiment), as shown in the following figure: \u003c\/p\u003e\n\u003cp\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231024\/82ba30e1290149eaad89ae57b5e604e3.png\" alt=\"\" width=\"346\" height=\"164\"\u003e\u003c\/p\u003e\n\u003cp\u003e4 Sample preparation \u003cbr\u003e Return the sample to room temperature and mix the sample before loading; If users need to dilute samples or high-concentration standards in the box, they can use the sample diluent in the box for dilution; If it is a cell sample, it is recommended to press before testing  3000rpm\/min  Rotational speed, centrifugation  5min , take the supernatant for detection. \u003cbr\u003e\u003cstrong\u003e Two, \u003c\/strong\u003e\u003cstrong\u003e Operation Process  \u003cbr\u003e\u003c\/strong\u003e1 Operation diagram \u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231024\/508efb921ac843ebb78e6c674b05a349.png\" alt=\"\" width=\"314\" height=\"406\"\u003e\u003cbr\u003e2 Detailed operation steps \u003cbr\u003e （ 1 ) Return each component of the kit to room temperature  30min , take out the enzyme-labeled slats required for the test from the aluminum foil bag that has been equilibrated to room temperature, mark the sequence of slats with a marker (it is recommended to perform double-hole determination), seal the remaining slats with a sealing film and put them back into the aluminum foil bag, seal them well, and place them in  2-8℃ Save. \u003cbr\u003e (Note: The slats are easy to fall off during the subsequent clapping process, so be careful to mark them) \u003cbr\u003e （ 2 ) Sample incubation: added to each well  50µL  Standard \u003cstrong\u003e\/\u003c\/strong\u003e Blank (sample dilution) \u003cstrong\u003e\/\u003c\/strong\u003e Sample, then added  50µL  Antibody working solution, sealed with plate sealing membrane, and then placed in  37℃ Static incubation in the dark  30min 。 \u003cbr\u003e (Note: The standard product must be added first. If the antibody is added first, it will directly react with the antigen on the plate; if the plate is not sealed or the plate is not sealed completely during the incubation process, the reaction solution will evaporate, resulting in errors in the experiment; Try to avoid light irradiation during all incubation processes) \u003cbr\u003e （ 3 ) Plate washing: After incubation, carefully peel off the plate sealing film, discard the liquid in the wells, and use the wash solution ( 1× ) wash plate  3  Times ( 250µL\/ Well), pat dry the residual liquid in the sample well. \u003cbr\u003e (Note: If hand washing plate is used, add washing solution ( 1× ), it is necessary to add liquid in the air, and it is best not to touch the inner wall of the hole with the gun tip; Each addition of the lotion ( 1× ) after standing  30s  And vibrate slightly; When patting dry, pay attention to changing absorbent paper each time or patting dry on clean areas of the paper) \u003cbr\u003e （ 4 ) Enzyme-labeled secondary antibody incubation: adding enzyme-labeled secondary antibody to each well, 100µL\/ Hole, seal the plate with a plate sealing film, and then place  37℃ Static incubation  30min 。 \u003cbr\u003e （ 5 ) Plate washing: the method is the same as the step   （ 3 ）。 \u003cbr\u003e （ 6 ) Color development: press the pre-configured color development solution  100µL\/ The holes are added into an enzyme label plate, and the plate is sealed with a plate sealing membrane, 37℃ Static incubation in the dark  15min 。 \u003cbr\u003e （ 7 ) Termination: add termination solution, 100µL\/ The hole can be read after the color is evenly developed. \u003cbr\u003e (Note: It is recommended to set in the microplate reader reading program  5-10s  Shock) \u003cbr\u003e （ 8 ) Reading: Place the microplate in the microplate reader and set the wavelength to dual wavelength  450\/630nm , read the absorbance value, and the determination should be terminated after  20min  Within completion. \u003cbr\u003e\u003cstrong\u003e Three, \u003c\/strong\u003e\u003cstrong\u003e Data processing  \u003cbr\u003e\u003c\/strong\u003e1 Calculation of absorbance value \u003cbr\u003e The absorbance value of each standard or sample is OD450nm-OD630nm 。 \u003cbr\u003e2 Calculation of percent absorbance \u003cbr\u003e The average absorbance value of each standard or sample (double well) divided by 0ng\/mL Of the standard   The average absorbance value is multiplied by 100% , that is, the percentage absorbance: \u003cbr\u003e Percent absorbance ( % ）＝ B\u003cstrong\u003e\/\u003c\/strong\u003eB0×100%\u003cbr\u003eB : Average absorbance value of standard or sample \u003cbr\u003eB0 ： 0ng\/mL Average absorbance values of standards \u003cbr\u003e3 Drawing and calculation of standard curve \u003cbr\u003e Take the percent absorbance value of the standard as ordinate Y The standard concentration value is the abscissa X , painted   Four parameters are recommended to prepare standard curves Logistic Mathematical model fitting equation: \u003cbr\u003eY=((A-D)\/(1+(X\/C)^B))+D\u003cbr\u003e Substitute the percentage absorbance value of the sample into the standard curve, read out the corresponding concentration value of the sample, and multiply it by its corresponding dilution factor to obtain the actual concentration of gentamicin in the sample. \u003cbr\u003e (Note: ① Per Chinese Pharmacopoeia \u0026lt;9012\u0026gt; 4. Relevant requirements in ligand binding analysis, four parameters are recommended for the fitting equation Logistic Mathematical models. ② Try to use the recommended dual-wavelength correction method, using 630nm Wavelength is corrected, OD450-OD630 Is the corrected OD The value can be directly used for calculation, or it can be blank corrected according to the data quality. If there is no dual wavelength microplate reader, read OD450 After the data, the data quality should be judged first, and then blank correction should be performed.) \u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231024\/899addb5e28e49b3aad4ce4cf1a99b89.png\" alt=\"\" width=\"375\" height=\"209\"\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis kit uses indirect competition ELISA method to determine trace residues of gentamicin in samples. The microwell strip is pre-coated with conjugated antigen, and the residual gentamicin in the sample competes with the conjugated antigen pre-coated on the microwell strip for anti-gentamicin antibody. The enzyme-labeled secondary antibody is added, and then the color is developed by adding TMB substrate. Use a microplate reader to measure the absorbance (OD value) at the wavelength of 450nm\/630nm, and the absorbance is negatively correlated with the content of gentamicin in the sample.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e         Gentamicin is an alkaline compound extracted from the fermentation broth of the genus Monospore of Actinomyceaceae. It is a commonly used aminoglycoside antibiotic and widely used in culture medium preparation. Clinically, it is mainly used to treat bacterial infections. Gentamicin is neurotoxic, nephrotoxic and ototoxic. Its residues in animal food and biological drugs will affect human health and even cause allergic reactions. European and American countries and China all require its limited use. \u003cbr\u003e         Gentamicin quantitative detection kit uses indirect competition ELISA Methods, the pre-coated gentamicin antigen on the microwell strip competed with the residual gentamicin in the sample to bind enzyme-labeled gentamicin monoclonal antibody, then the enzyme-labeled secondary antibody was added, and then by adding TMB The substrate developed color, and the absorbance value was detected by a microplate reader. The absorbance value was negatively correlated with the content of gentamicin in the sample. The operation time of this kit is only about one hour, and the linear range is 0.1ng\/mL-10ng\/mL 。 \u003cbr\u003e         When doing the linearity verification of gentamicin raw materials, it is recommended to dilute gentamicin raw materials within the linear range of the kit before determination, and dilute them according to a certain proportion to at least  5  The detected concentration is linearly correlated with the dilution factor.  \u003cbr\u003e         Specificity should be verified for other substances present in the process. It is recommended to use high-concentration and low-concentration quality control samples, and add increasing concentrations of relevant interfering substances for specific investigation. The matrix to which no analyte is added should also be measured at the same time. The accuracy of the controls should be within a range and the measured values of the matrix without the addition of analyte should be below the lower limit of quantification.  \u003cbr\u003e\u003cstrong\u003e Product Features: \u003c\/strong\u003e\u003cbr\u003e Strong specificity: no cross-reaction with penicillin, streptomycin sulfate, kanamycin, ampicillin \u003cbr\u003e High sensitivity: the kit sensitivity is 0.1ng\/mL\u003cbr\u003e Convenient and efficient: using the two-step method, just 1 The results can be obtained in about hours \u003cbr\u003e Excellent performance: good calibration curve, high accuracy and good repeatability \u003cbr\u003e\u003cstrong\u003e Product performance indicators: \u003c\/strong\u003e\u003cbr\u003e Limit of detection: \u0026lt; 0.1 ng\/mL ；  \u003cbr\u003e Limit of quantification: 0.1 ng\/mL \u003cbr\u003e Linearity Range: 0.1-10 ng\/mL \u003cbr\u003e Accuracy (spiked recovery): 70%-130% \u003cbr\u003e Accuracy (measurement deviation): ≤15% \u003cbr\u003e Repeatability (intra-batch difference): ≤15% \u003cstrong\u003e\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e Specificity: \u003c\/strong\u003e\u003cbr\u003e\u003ctable style=\"width: 327px;\" border=\"1\" width=\"345\" cellspacing=\"0\" cellpadding=\"0\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 169px;\" width=\"221\" height=\"28\"\u003e\u003cspan class=\"font1\"\u003e Antibiotics \u003c\/span\u003e\u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"text-align: center; width: 128px;\" width=\"297\"\u003e Cross-reactivity rate \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"text-align: center; width: 169px;\" height=\"28\"\u003e Gentamicin \u003c\/td\u003e\n\u003ctd class=\"et7\" style=\"text-align: center; width: 128px;\"\u003e100%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et3\" style=\"text-align: center; width: 169px;\" height=\"28\"\u003e Streptomycin sulfate \u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"text-align: center; width: 128px;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et4\" style=\"text-align: center; width: 169px;\" height=\"28\"\u003e Kanamycin \u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"text-align: center; width: 128px;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et3\" style=\"text-align: center; width: 169px;\" height=\"28\"\u003e Penicillin \u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"text-align: center; width: 128px;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et5\" style=\"text-align: center; width: 169px;\" height=\"28\"\u003e Ampicillin \u003c\/td\u003e\n\u003ctd class=\"et6\" style=\"text-align: center; width: 128px;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003ctable style=\"width: 69.1196%;\" border=\"1\" width=\"69.1196%\" cellspacing=\"0\" cellpadding=\"0\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" width=\"96\" height=\"28\"\u003e serial number \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\" width=\"252\"\u003e Name \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\" width=\"139\"\u003e Specifications \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\" width=\"274\"\u003e Save \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e1\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Enzyme plate \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e8×12  Strip \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃ Protect from light \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e2\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Standard ( 100 ng\/mL ） \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃ Protect from light \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e3\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Antibody working solution \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e7mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e4\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Enzyme-labeled secondary antibody \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e12mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e5\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Sample dilution \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e30mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e6\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Lotion ( 20× ） \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e30mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e7\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Chromogenic liquid  A\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e8mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃ Protect from light \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e8\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Chromogenic liquid  B\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e8mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃ Protect from light \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e9\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Stop liquid \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e15mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd class=\"et2\" style=\"width: 11.2088%; text-align: center;\" height=\"28\"\u003e10\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.4819%; text-align: center;\"\u003e Sealing film \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.2947%; text-align: center;\"\u003e3 Zhang \u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 22.38%; text-align: center;\"\u003e-\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. All components in the kit must be restored to room temperature (20-25 ℃) before use.  \u003cbr\u003e2. Mix all components thoroughly before use. The standard product needs to be shortly centrifuged for 5 seconds to concentrate all the liquid on the tube wall and lid at the bottom of the tube; All reagents were returned to 2-8 °C immediately after use.  \u003cbr\u003e3. The kit must be used within the validity period, and the corresponding standard curve must be prepared for each test. It is not recommended to use different batches of related reagents in mixed batches.  \u003cbr\u003e4. When adding liquid to the microplate, be careful not to touch the bottom of the microplate to prevent damage to the coating layer. Replace the sample loading tank and suction tip in time between different samples and steps to avoid cross-contamination.  \u003cbr\u003e5. When patting the slats dry after washing, be careful to prevent the slats from falling off, and be careful not to reuse the sealing film.  \u003cbr\u003e6. High concentration may produce black flocs during color development, which is a normal phenomenon, and the slight degree will not affect the final reading result.  \u003cbr\u003e7. When reading, pay attention to check whether the detection wavelength and fitting equation are correct.  \u003cbr\u003e8. Only by strictly abiding by the operation methods of the instructions and using all the reagents supporting this kit can the best detection effect be guaranteed.  \u003cbr\u003e9. Differences in test results can be caused by a variety of factors, including the operation of the experimenter, the use of the pipette, the plate washing technique, the reaction time or temperature, the storage of the kit, etc.  \u003cbr\u003e10. The company is only responsible for the kit itself, and is not responsible for the sample consumption caused by the use of the kit. Users are requested to fully consider the possible usage of samples and reserve sufficient samples before use.  \u003cbr\u003e11. The termination solution in this kit is acid solution, so special attention should be paid to operation.  \u003cbr\u003e12. Operators should wear personal protective equipment, such as laboratory coats, gloves, masks and goggles.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eThe kit should be stored at 2-8 ℃, protected from light, and the shelf life is 12 months. Note that the unused kit after opening should still be stored at 2-8 ℃ in the dark.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293314981963,"sku":"abs590011-96T","price":663.0,"currency_code":"USD","in_stock":true}]},{"product_id":"collagenase-type-i-residue-detection-kit","title":"Collagenase Type I Tesidue Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e1. Preparation work:\u003c\/strong\u003e\u003cbr\u003e1. Kit preparation\u003cbr\u003eEach component of the kit was left to equilibrate at room temperature for 30 minutes before operation was started.  \u003cbr\u003e2. Required consumables and equipment (Note: You need to bring your own)\u003cbr\u003e(1) Microplate reader, constant temperature oscillator (or constant temperature incubator), plate washer (the plate washing method is hand washing or not), vortex oscillator, timer\u003cbr\u003e(2) High precision pipettes and disposable tips (0.5-10 µ L, 10-100 µ L, 30-300 µ L, 100-1000 µ L)\u003cbr\u003e(3) Deionized water\u003cbr\u003e(4) Absorbent paper, EP tube, disposable gloves\u003cbr\u003e3. Reagent preparation (Note: prepare according to the requirements according to the components provided in the kit)\u003cbr\u003e(1) Wash solution (1 ×): Take the wash solution (20 ×) and add deionized water to dilute it 20 times for later use. For example, take 10mL of wash solution (20 ×) and add 190mL of deionized water to mix well.\u003cbr\u003eNote: If crystals are formed in the wash solution (20 ×), shake gently in room temperature or 37 ℃ water bath, and then dilute after the crystals are completely dissolved)\u003cbr\u003e(2) Preparation of enzyme-labeled antibody solution: Dilute enzyme-labeled antibody (10 ×) 10 times with diluent to prepare enzyme-labeled antibody solution (1 ×). For example: 100μL enzyme-labeled antibody is added to 900μL diluent.\u003cbr\u003e(3) Preparation of color development solution: Mix color development solution A, color development solution B and other volumes, mix well and place it in the dark from light (Note: The time cannot be left for too long, generally prepare it 10min before use. If the color development solution has turned blue after mixing, please do not use it).  \u003cbr\u003e4. Preparation of standards\u003cbr\u003e(Note: It is prepared according to the requirements according to the standard provided by the kit. In order to further ensure the accuracy of the results, you can also use collagenase from specific sources actually used in the production process to establish the standard curve by yourself)\u003cbr\u003eDilute the standard (1 μg\/mL) to 20 ng\/mL with a diluent and then prepare the standard by serial dilution (dilution factor: 2 times) as shown in the following figure:\u003cbr\u003e\u003cimg src=\"http:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin_new_bak-english\/20260630\/b89816196a3e4540addb17a4e9763012.jpg\" alt=\"\" width=\"500\" height=\"230\"\u003e\u003cbr\u003eNote: In order to ensure the validity of the experimental results, a freshly prepared standard solution is used for each experiment.  \u003cbr\u003e5. Sample preparation\u003cbr\u003eReturn the sample to room temperature and mix well before adding the sample (the sample should be a homogeneous solution, and if there is precipitation, centrifuge the supernatant); If users need to dilute samples or high-concentration standards in the box, they can use the diluent in the box for dilution.\u003cbr\u003e\u003cstrong\u003e2. Operation process\u003c\/strong\u003e\u003cbr\u003e1. Detailed operation steps\u003cbr\u003eAll operations are performed at room temperature and repeat assays are recommended for all spiked wells.  \u003cbr\u003e(1) Return each component of the kit to room temperature for 30min, take out the slats required for the test from the aluminum foil bag that has been equilibrated to room temperature, mark the sequence of the slats with a marker, seal the remaining slats with a sealing film and put them back in the aluminum foil bag, seal them well, and store them at 2-8 ℃.  \u003cbr\u003e(Note: The slats are easy to fall off during the washing process, so be careful to mark them.)\u003cbr\u003e(2) Sample incubation: Add the diluted standard substance and the sample to be tested to the enzyme label plate (recommended addition sequence: standard well, blank well, sample well, standard substance is added according to the concentration gradient), 100 µ L\/well, seal the plate with a plate sealing membrane, and then place it at 37 ℃ for incubation for 1h.  \u003cbr\u003e(Note: The sample addition time should be controlled within 10min to avoid drift over time. If the plate is not sealed or the plate is incomplete during incubation, the reaction solution will evaporate, resulting in errors in the experiment.)\u003cbr\u003e(3) Plate washing: After incubation, carefully peel off the sealing film, discard the liquid in the wells, wash the plates three times (250 µ L\/well) with washing solution (1 ×), and pat dry the residual liquid in the sample wells. (If the plate washing method is hand washing, it can be allowed to stand for 1min after adding washing solution (1 ×); if the plate washing machine is used to wash the plates, it can be slightly shaken for 5s after adding washing solution.)\u003cbr\u003e(4) Incubation of enzyme-labeled antibody solution: Add enzyme-labeled antibody solution to each well, 100 µ L\/well, seal the plate with a plate sealing membrane, and then place it at 37 °C for 1h.  \u003cbr\u003e(Note: Before adding liquid after each plate washing, check whether the slats are fixed to prevent liquid splashing caused by fixing the slats after adding liquid.)\u003cbr\u003e(5) Plate washing: same as step 3).  \u003cbr\u003e(6) Color development: Add the pre-prepared color development solution to the enzyme labeled plate, 100 µ L\/well, seal the plate with a sealing membrane, and incubate at 37 °C in the dark for 20 minutes.  \u003cbr\u003e(7) Termination: Add the stop solution, 100 µ L\/well, and the reading can be made after the color is uniform (Note: Generally, the reading can be completed within 20 minutes after adding the stop solution).  \u003cbr\u003e(8) Reading: Put the microplate into the microplate reader, set the wavelength to dual wavelength 450\/630 nm, and read the absorbance value (Note: It is recommended to set a 5-10s shock in the microplate reader reading program).\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e3. Data processing:\u003c\/strong\u003e\u003cbr\u003e1. Calculation of absorbance value\u003cbr\u003eThe light absorption calibration values for each standard or sample are:\u003cbr\u003eStandard\/sample well absorbance values (OD\u003csub\u003e450nm\u003c\/sub\u003e-OD\u003csub\u003e630nm\u003c\/sub\u003e)-Blank control well absorbance value (OD\u003csub\u003e450nm\u003c\/sub\u003e-OD\u003csub\u003e630nm\u003c\/sub\u003e） \u003cbr\u003e2. Draw the standard curve with the standard concentration calibration value as the abscissa (X) and the standard light absorption calibration value as the ordinate (Y). It is recommended to use a four-parameter Logistic mathematical model to fit the equation:\u003cbr\u003eY = ((A-D)\/(1 + (X\/C) ^ B)) + D\u003cbr\u003eLight absorption calibration value [standard\/sample well absorbance value (OD\u003csub\u003e450nm\u003c\/sub\u003e-OD\u003csub\u003e630nm\u003c\/sub\u003e)-Blank control well absorbance value (OD\u003csub\u003e450nm\u003c\/sub\u003e-OD\u003csub\u003e630nm\u003c\/sub\u003e)] Substitute into the formula to calculate the content of collagenase in the sample.  \u003cbr\u003e3. If the OD value of the sample to be tested exceeds the OD value of the highest point of the standard curve, the sample needs to be diluted and re-measured\u003cbr\u003eThe following standard curve is for reference only, and the standard curve drawn by the same experimental standard shall prevail.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis kit uses double antibody sandwich enzyme-linked immunosorbent assay to determine the trace residue of Collagenase Type I (hereinafter referred to as collagenase) derived from Clostridium histolyticum in samples. Coating 96-well plates with capture antibodies to make solid phase antibodies, then adding standard and test samples, and then adding horseradish peroxidase (HRP) labeled labeled antibodies to form a solid phase antibody-collagenase-labeled antibody sandwich conjugate. Its absorbance (OD value) was measured at wavelengths of 450 nm and 630 nm, where 630 nm is the corrected wavelength. Results Calculate the standard\/sample well absorbance value (OD450nm-OD630nm)-blank control well absorbance value (OD450nm-OD630nm), and calculate the content of collagenase in the sample to be tested through the standard curve (it is recommended to use the four-parameter Logistic mathematical model fitting equation).\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCollagenase can be used to separate various tissues into single cells in vitro, and is suitable for single cell dissociation of organs and tissues for various scientific research purposes. The regulations related to the production of biological products clarify the necessity of detection of cell culture-related materials, so the detection of collagenase residues has attracted more and more attention.  \u003cbr\u003eThis product uses a double antibody sandwich enzyme-linked immunosorbent assay based on polyclonal antibody, developed for collagenase mixtures obtained from Clostridium histolyticum, which can quickly and effectively detect collagenase residues in various samples.\u003cbr\u003e\u003cstrong\u003eProduct Features:\u003cbr\u003e\u003c\/strong\u003eAccurate detection: Polyclonal antibodies prepared against collagenase mixtures can perform effective detection\u003cbr\u003eStrong specificity: no cross-reaction with other non-target proteins commonly used in cell culture\u003cbr\u003eExcellent performance: detection limit as low as 0.1 ng\/mL, high accuracy and good repeatability\u003cbr\u003eWide applicability: You can use collagenase from specific sources actually used in the production process to establish a standard curve by yourself to further ensure the accuracy of the results.\u003cbr\u003e\u003cstrong\u003eProduct performance indicators:\u003cbr\u003e\u003c\/strong\u003eLinearity range: 0.625-20ng\/mL\u003cbr\u003eLimit of quantitation: 0.625 ng\/mL\u003cbr\u003eLimit of detection: ≤ 0. 5 ng\/mL\u003cbr\u003eAccuracy (spiked recovery): 80%-120%\u003cbr\u003eAccuracy (measurement deviation): ≤ 15%\u003cbr\u003eRepeatability (intra-batch difference): ≤ 10%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003ctable style=\"height: 254px; width: 69.1196%;\" border=\"1\" width=\"69.1196%\" cellspacing=\"0\" cellpadding=\"0\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 28px;\"\u003e\n\u003ctd class=\"et2\" style=\"width: 11.9693%; text-align: center; height: 28px;\" width=\"96\" height=\"28\"\u003eserial number\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.672%; text-align: center; height: 28px;\" width=\"252\"\u003eName\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.865%; text-align: center; height: 28px;\" width=\"139\"\u003eSpecifications\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.8591%; text-align: center; height: 28px;\" width=\"274\"\u003eSave\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 28px;\"\u003e\n\u003ctd class=\"et2\" style=\"width: 11.9693%; text-align: center; height: 28px;\" height=\"28\"\u003e1\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.672%; text-align: center; height: 28px;\"\u003eEnzyme plate\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.865%; text-align: center; height: 28px;\"\u003e8 × 12 bars\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.8591%; text-align: center; height: 28px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px;\"\u003e\n\u003ctd class=\"et2\" style=\"width: 11.9693%; text-align: center; height: 10px;\" height=\"28\"\u003e2\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.672%; text-align: center; height: 10px;\"\u003eStandard (1 µ g\/mL)\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.865%; text-align: center; height: 10px;\"\u003e\u003cp\u003e300 µ L\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.8591%; text-align: center; height: 10px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.9693%; text-align: center; height: 22px;\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 35.672%; text-align: center; height: 22px;\"\u003eSample dilution\u003c\/td\u003e\n\u003ctd style=\"width: 20.865%; text-align: center; height: 22px;\"\u003e60mL\u003c\/td\u003e\n\u003ctd style=\"width: 20.8591%; text-align: center; height: 22px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.9693%; text-align: center; height: 22px;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 35.672%; text-align: center; height: 22px;\"\u003eEnzyme-labeled antibody (10 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 20.865%; text-align: center; height: 22px;\"\u003e1.5 mL\u003c\/td\u003e\n\u003ctd style=\"width: 20.8591%; text-align: center; height: 22px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.9693%; text-align: center; height: 22px;\"\u003e5\u003c\/td\u003e\n\u003ctd style=\"width: 35.672%; text-align: center; height: 22px;\"\u003eLotion (20 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 20.865%; text-align: center; height: 22px;\"\u003e30mL\u003c\/td\u003e\n\u003ctd style=\"width: 20.8591%; text-align: center; height: 22px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 28px;\"\u003e\n\u003ctd class=\"et2\" style=\"width: 11.9693%; text-align: center; height: 28px;\" height=\"28\"\u003e6\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.672%; text-align: center; height: 28px;\"\u003eChromogenic solution A\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.865%; text-align: center; height: 28px;\"\u003e8mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.8591%; text-align: center; height: 28px;\"\u003e2-8 ℃ protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 28px;\"\u003e\n\u003ctd class=\"et2\" style=\"width: 11.9693%; text-align: center; height: 28px;\" height=\"28\"\u003e7\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 35.672%; text-align: center; height: 28px;\"\u003eChromogenic solution B\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.865%; text-align: center; height: 28px;\"\u003e8mL\u003c\/td\u003e\n\u003ctd class=\"et2\" style=\"width: 20.8591%; text-align: center; height: 28px;\"\u003e2-8 ℃ protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.9693%; text-align: center; height: 22px;\"\u003e8\u003c\/td\u003e\n\u003ctd style=\"width: 35.672%; text-align: center; height: 22px;\"\u003eStop liquid\u003c\/td\u003e\n\u003ctd style=\"width: 20.865%; text-align: center; height: 22px;\"\u003e15mL\u003c\/td\u003e\n\u003ctd style=\"width: 20.8591%; text-align: center; height: 22px;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.9693%; text-align: center; height: 22px;\"\u003e9\u003c\/td\u003e\n\u003ctd style=\"width: 35.672%; text-align: center; height: 22px;\"\u003eSealing film\u003c\/td\u003e\n\u003ctd style=\"width: 20.865%; text-align: center; height: 22px;\"\u003e3 sheets\u003c\/td\u003e\n\u003ctd style=\"width: 20.8591%; text-align: center; height: 22px;\"\u003e-\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBackground\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCollagenase is a collagenase mixture obtained from Clostridium histolyticum, which can be used to separate a variety of tissues into single cells in vitro, and is suitable for single cell dissociation of organs and tissues for various scientific purposes. Collagenase is widely used in the isolation and acquisition of mesenchymal stem cells, 3D and organoid culture, single cell sequencing, tumor research and other fields. The bacterial collagenase commercially available on the market is mainly derived from Clostridium histolyticum. It is a crude enzyme extract that not only contains collagenase (clostridiopeptidase A), but also can degrade natural collagen and reticular fibers. It also contains some other proteases, polysaccharide enzymes, lipases, etc. According to the difference of collagenase activity, it can be divided into collagenase type I, type II, type III, type IV and type V, which can act on different types of tissue dissociation. Bacterial collagenases differ from vertebrate collagenases in that they have a wider substrate singularity. Unlike animal collagenase, which only decomposes the natural three-dimensional helical structure of collagen, bacterial collagenase is unique in that it can degrade both water-insoluble natural collagen and water-soluble denatured proteins. Bacterial collagenase can degrade almost all collagen types, and there are many kinds of divisions in the three-dimensional helical region. Collagenase is an endopeptidase, which can specifically hydrolyze the three-dimensional helical structure of natural collagen under physiological temperature and PH conditions. Collagen is the main fibrous component of animal extracellular connective tissue. This function of collagenase is mainly due to its ability to specifically recognize the Pro-X-Gly-Pro sequence and cleave the peptide bond between the neutral amino acid (X) and glycine (Gly) of this sequence, which is present in collagen with high frequency. Collagenase is also the only protease that can degrade natural collagen fibers with triple helices widely existing in connective tissue. Collagenase is also efficient enough to hydrolyze other proteins, polysaccharides and lipids within the extracellular matrix of connective tissue and epithelial tissue, making collagenase products well suited for tissue dissociation. In the treated biological products, there may be trace collagenase residues, which will have a certain impact on the subsequent application of biological products. The residue of collagenase in biological products is one of the important indexes to measure the quality of biological products.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. All components in the kit must be restored to room temperature (20-25 ℃) before use.  \u003cbr\u003e2. Each component should be thoroughly mixed before use to ensure the uniformity of the reagents. The standard product should be centrifuged briefly for 5 seconds. All the liquid on the tube wall and lid should be concentrated at the bottom of the tube. Immediately after use, all the reagents should be put back to 2-8 ℃.  \u003cbr\u003e3. The kit must be used within the validity period, and the corresponding standard curve must be re-prepared for each test. It is not recommended to mix different standard curves, and it is not recommended to mix different batches of related reagents.  \u003cbr\u003e4. When adding liquid to the microplate, be careful not to touch the bottom of the microplate to prevent damage to the coating layer. Change the sample loading tank and tip in time between different samples to avoid cross-contamination.\u003cbr\u003e5. When patting the slats dry after washing, be careful to prevent the slats from falling off, and be careful not to reuse the sealing film.  \u003cbr\u003e6. High concentration may produce black flocs during color development. It is recommended to dilute the sample for retest.  \u003cbr\u003e7. When reading, pay attention to check whether the detection wavelength and fitting equation are correct.  \u003cbr\u003e8. Only by strictly abiding by the operation methods of the instructions and using all the reagents supporting this kit can the best detection effect be guaranteed.  \u003cbr\u003e9. The kit uses accurate quantification of high-purity and high-activity collagenase as the standard product, which is suitable for most type I collagenases derived from Clostridium histolyticum. It can directly detect the residual levels of collagenase from different sources and can be used for more accurate detection of Merck Sigma-Aldrich collagenase. To further guarantee the accuracy of the results, it is also possible to self-establish standard curves using specific sources of collagenase actually used in the manufacturing process.  \u003cbr\u003e10. Differences in test results can be caused by many factors, including the operation of the experimenter, the use of the pipette, the plate washing technique, the reaction time or temperature, the storage of the kit, etc.  \u003cbr\u003e11. The company is only responsible for the kit itself, and is not responsible for the sample consumption caused by the use of the kit. Users are requested to fully consider the possible usage of samples and reserve sufficient samples before use.  \u003cbr\u003e12. The termination solution in this kit is acid solution, so special attention should be paid to operation.  \u003cbr\u003e13. For safety reasons, operators should wear personal protective equipment, such as lab coats, gloves, masks and goggles.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Collagenase Type I quantitative detection kit (enzyme-linked immunosorbent assay) produced by our company is a highly sensitive collagenase detection kit, which is developed for the collagenase mixture obtained from Clostridium histolyticum, and can sensitively, specifically and accurately detect the residue of type I collagenase in intermediate products, semi-finished products and finished products of various tissue dissociation samples. The kit uses accurate quantification of high-purity and high-activity collagenase as the standard product, which is suitable for most type I collagenases derived from Clostridium histolyticum, and can directly detect the residual levels of collagenase from different sources. The standard can be used for more accurate detection of Merck Sigma-Aldrich collagenase type I. To further guarantee the accuracy of the results, it is also possible to self-establish standard curves using specific sources of collagenase actually used in the manufacturing process.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe kit should be stored at 2-8 ℃, protected from light, and the shelf life is 12 months. Note that the unused kit after opening should still be stored at 2-8 ℃ in the dark.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293315047499,"sku":"abs590012-96T","price":1375.0,"currency_code":"USD","in_stock":true}]},{"product_id":"limulidae-kit","title":"Dynamic chromogenic endotoxin detection recombinant tachypleus amebocyte lysate Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003ematerials required: \u003c\/strong\u003eendotoxin working standard, water for bacterial endotoxin test, pyrogen removal test tube, pyrogen Removal Tip, pyrogen removal 96 well microplate (or detachable strip), vortex mixer, pipette, test tube rack, dynamic photometric instrument with incubation system and supporting software\u003cbr\u003e\u003cbr\u003e1The setting of the dynamic photometric instrument \u003cbr\u003epreheat the instrument and set the incubation temperature to 37º C. Set up templates and procedures. Set the plate reading wavelength to 405 nm, and set the kinetic parameters: read the plate for 60 minutes, once every 30 seconds\u003cbr\u003e2Preparation of endotoxin standard solution \u003cbr\u003ea Concentration of dynamic color development method: 10, 1, 0.1, 0.01 eu\/ml or 5, 0.5, 0.05, 0.005 eu\/ml\u003cbr\u003eb Concentration by rate method: 0.1, 0.05, 0.025, 0.0125, 0.00625 eu\/ml\u003cbr\u003e2.1Take one endotoxin working standard, scratch it along the neck of the ampoule with a grinding wheel, open it, add an appropriate amount of water for bacterial endotoxin test, and place it on the vortex mixer to shake vigorously for 2 minutes\u003cbr\u003e2.2Further dilute the above endotoxin solution with water for bacterial endotoxin test to the required concentration, and shake vigorously on the vortex mixer for 1 minute at each dilution step. The diluted endotoxin solution shall be left standing for more than 10 minutes. Before use, it shall be vigorously shaken on the vortex mixer for 1 minute. The endotoxin solution placed for more than 4 hours shall be discarded\u003cbr\u003e3The negative control was water for bacterial endotoxin test\u003cbr\u003e4Configuration of recombinant tachypleus amebocyte lysate: pipette 3 ml of the complex solution into the lyophilized recombinant tachypleus amebocyte lysate, gently shake or blow with a pyrogen Removal Tip to completely mix the recombinant tachypleus amebocyte lysate. The dissolved recombinant tachypleus amebocyte lysate should be used within 20 minutes\u003cbr\u003e5In the experimental operation, remove the pyrogen microplate, and add 100 UL of bacterial endotoxin test water, endotoxin standard solution, or test article into each well. Add 100 UL of recombinant tachypleus amebocyte lysate per well with a pipette or multi-channel pipette (\u003cstrong\u003eto avoid bubbles \u003c\/strong\u003e), mix with medium speed vortex for 15 seconds, and put it into the microplate reader to read according to the set parameters\u003cbr\u003e6Data processing \u003cbr\u003e6.1Threshold method: \u003cbr\u003eset the threshold to 0.1 (which can be adjusted according to the result), paste the data into the calculation table according to the specification, and the calculation table will automatically give its start time (CT). According to the requirements of the calculation table, fill in the concentration value of the working calibrator, and automatically establish the standard curve: lgct = B LGC + A, where CT is the start-up time, C is the concentration of endotoxin, B is the linear slope, and a is the y-axis intercept. The experiment is valid only when the experimental data meet the following three conditions at the same time: ① the concentration point of the standard curve ≥ 4, correlation coefficient of standard curve | R | ≥ 0.980, ② the T value of the lowest point of the standard curve is less than that of the negative control, ③ the average value of the parallel tubes of the test article is within the range of the standard curve\u003cbr\u003e6.2Rate method: \u003cbr\u003efor samples with concentration below 0.1 eu\/ml, the rate method can also be used, and the reading time is 30 minutes. The rate method calculation table is used for data processing, and the reaction time parameters can be adjusted within 900-1800 seconds\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003einterference test of test article \u003c\/strong\u003e\u003cbr\u003e① when endotoxin test method is established for new drugs or varieties without endotoxin test items, interference test must be carried out\u003cbr\u003e② when the source, formula and production process of tachypleus amebocyte lysate and test article are changed, or any change that may affect the test results occurs in the test environment, the interference test must be repeated\u003cbr\u003e③ when there may be interfering substances of Limulus amebocyte lysate test in the test article, the interference test must be carried out\u003cbr\u003ethe steps are as follows: \u003cbr\u003e1Select the midpoint of the standard curve or an endotoxin concentration close to the midpoint (set as λ m) as the endotoxin concentration added in the test article interference test. If the standard curve is 10, 1, 0.1, 0.01eu\/ml series, 0.1eu\/ml endotoxin concentration can be prepared with the test article as the positive control of the test article\u003cbr\u003e2The concentration of the test solution is λ M endotoxin solution (i.e., the positive control of the test sample containing λ m endotoxin), and the endotoxin concentration of the solution is measured, which is called CS\u003cbr\u003e3Measure the endotoxin concentration of the test solution without exogenous endotoxin, which is called C0\u003cbr\u003e4Calculate the recovery rate r = (cs– C0) \/λ M × 100%\u003cbr\u003e5When R is between 50% and 200%, it is considered that the test solution has no obvious interference effect under this test condition\u003cbr\u003e6When R is beyond 50% - 200%, serial dilution or other treatment shall be carried out on the test article to eliminate interference. Repeat steps 2-4 for each diluted solution until the recovery rate of endotoxin R is between 50% - 200%. The dilution multiple with the recovery r closest to 100% was selected for endotoxin detection. (refer to bacterial endotoxin test method in Pharmacopoeia of the People'sRepublic ofChina, 2020 Edition)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe recombinant horseshoe crab reagent contains three recombinant proteins: factor C, factor B, and prothrombin. The reaction principle is shown in Figure 1. Bacterial endotoxins activate factor C, causing a series of enzymatic reactions, activating coagulase to form coagulase, which decomposes the artificially synthesized chromogenic matrix into peptides and yellow p-nitroaniline (pNA), λ Max=405). The higher the concentration of endotoxin, the faster the release of pNA and the faster the absorbance change.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eEndotoxin testing\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eRecombinant tachypleus amebocyte lysate is an alternative to pharmacopoeia testing, which can be used for the final product testing of human injected drugs (including biological products), animal injected drugs and medical equipment products. Guidance on the validation of alternative test methods can be found at USP \u0026lt;1223\u0026gt;And \u0026lt;1225\u0026gt;Found in. Such methods should prove to be equivalent or better than pharmacopoeial methods\u003cbr\u003ethe test method of this kit can also be used for quantitative detection of endotoxin in non pharmacopoeial substances (for example, raw materials including water and in-process monitoring), without method validation\u003cbr\u003erecombinant tachypleus amebocyte lysate is not suitable for detecting endotoxin in clinical samples, such as those in clinical samples for the diagnosis of human diseases (such as endotoxemia)\u003cbr\u003e\u003cstrong\u003eproduct features: \u003c\/strong\u003e\u003cbr\u003e1The traditional Limulus amebocyte lysate has no sensory substitution: the operation process is unchanged, the detection equipment is the same, and the analysis method is the same\u003cbr\u003e2No animal ingredients - no tachypleus amebocyte lysate blood required\u003cbr\u003e3The same cascade reaction process\u003cbr\u003e4With 1, 3-\u0026amp;beta-D glucan did not react. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTwo bottles of freeze-dried recombinant limulus lysate, two bottles of compound solution, and two bottles of endotoxin working standard\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. this kit is used for quantitative detection of bacterial endotoxin in vitro. It is strictly forbidden for reagents to enter the body in any way\u003cbr\u003e2All vessels in contact with reagents and test articles must be pyrogen free. Microbial contamination should be prevented during the test. The pyrogen in this manual refers to endotoxin concentration less than 0.005 eu\/ml\u003cbr\u003e3The pH value of the test article should be 6.0 – If it is beyond this range, it needs to be adjusted with pyrogen removal buffer, 0.1M sodium hydroxide or 0.1M hydrochloric acid\u003cbr\u003e4In the interference test of the test article, if there may be interference substances of the recombinant tachypleus amebocyte lysate test in the test article, the interference test must be carried out. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplication\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eQuantitative testing of endotoxin quantification for non pharmacopoeial items (such as raw materials including water and monitoring during the process)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e2-8° Cprotected from light, 2 years\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"1kit","offer_id":41518136623179,"sku":"abs50045-1kit","price":250.0,"currency_code":"USD","in_stock":true}]},{"product_id":"dsrna-elisa-kit-j2-based","title":"Double-stranded RNA (dsRNA) ELISA kit (J2 based)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eSee the instruction manual\u003cbr\u003eattached to the goods.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eForm\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eReagent test kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eSandwich elisa \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eDouble-stranded RNA(dsRNA) ELISA Kit(J2 based)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eJ2 monoclonal antibody coated Microplate, K2 detection antibody, dsRNA standard, HRP HRP-labeled secondary antibody, TMB color development solution, termination solution, plate sealing membrane, etc.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBackground\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eBased on the use of two double-stranded RNA (dsRNA) -specific monoclonal antibodies, the dsRNA detection kit can sensitively and selectively detect dsRNA molecules (greater than 30 to 40 bp) independent of their nucleotide composition and sequence. Detection with high specificity, in the presence of 1.000 to 10.000 times the other nucleic acid under the condition of excessive, can detect dsRNA in nucleic acid extraction. The assay was based on a sandwich ELISA using J2 (IgG2a) mouse monoclonal antibody against dsRNA as a capture antibody. Monoclonal antibody K2 (IgM) was used as the detection antibody. J2 anti-dsrna IgG2a monoclonal antibody has become the gold standard for dsRNA detection. It was originally used for the study of plant viruses, but since the seminal paper by Weber et al. In 2006, J2 was used to demonstrate that all plus-strand RNA viruses tested produced large amounts of dsRNA in infected cells, and this antibody has been widely used in a variety of systems, as described in more than 200 scientific publications. J2 can be used to detect dsRNA intermediates of a variety of viruses such as hepatitis C virus, dengue virus, rhinovirus, chikungunya virus, rabies virus, poliovirus, classical swine fever virus, Brome Mosaic virus and many more present in cultured cells and fixed paraffin embedded histological samples.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis product is only for research and in vitro testing, and is not suitable for diagnostic or therapeutic procedures involving humans or animals. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplication\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis kit can detect viral dsRNAs or large natural or synthetic dsRNAs from non viral sources in nucleic acid extracts, as well as detect the presence of unwanted dsRNA molecules in artificially synthesized (m) RNA preparations. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe whole kit was stored in -20º C lowerAfter the kit is thawed, it can be stored in 4º C for 5 daysFor long-term storage, it is recommended to store in – 20º Aliquot and freeze the antibody and dsRNA components under C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41518198882379,"sku":"abs590003-96T","price":917.0,"currency_code":"USD","in_stock":true}]},{"product_id":"bsa-elisa-lit","title":"BSA ELISA Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e 1. Preparation work: \u003c\/strong\u003e\u003cbr\u003e1 Kit preparation: Each component of the kit is placed at room temperature for equilibrium 30min After starting the operation.\u003cbr\u003e2 Consumables and equipment that need to be brought by yourself: microplate reader, constant temperature oscillator (or constant temperature incubator), plate washing machine (it is recommended not to cross-use with other items or directly use hand washing), vortex oscillator, timer, high-precision pipettes and disposable tips ( 0.5-10uL 、 10-100uL 、 30-300uL 、 100-1000uL ), sterile deionized water without heat source, absorbent paper, EP Tubes, disposable gloves.\u003cbr\u003e3 Reagent preparation: ( \u003cstrong\u003e Note: \u003c\/strong\u003e Try not to weigh in the laboratory BSA Powder or not left weighed BSA Electronic balance, clean the experimental table before the experiment, and prepare reagents according to the requirements of the current experiment)\u003cbr\u003e① Buffer （ 1× ) Preparation: take Buffer （ 20× ) Dilute with deionized water 20 Times for standby use, such as: take Buffer （ 20× ） 10mL Add DI water 190mL Mix well. ( \u003cstrong\u003e Note: \u003c\/strong\u003eBuffer （ 20× If crystals are formed in), it should be placed at room temperature or 37℃ Shake gently in water bath and wait until the crystals are completely dissolved before diluting)\u003cbr\u003e② Preparation of color developing solution: The color developing solution A Chromogenic Solution B Mix in equal volumes, mix well and place in the dark. ( \u003cstrong\u003e Note: \u003c\/strong\u003e The time cannot be left for too long, generally before use 10min Formulation. Do not use if the colored solution has turned blue after mixing)\u003cbr\u003e③ Enzyme-labeled antibody ( 1× ) Preparation: Take out the enzyme-labeled antibody and put it on the ice box, according to the experimental dosage ( 100uL\/ Hole), with Buffer （ 1× ) Dilute the enzyme-labeled antibody 500 Times is the enzyme-labeled antibody ( 1× ）。（ \u003cstrong\u003e Note: \u003c\/strong\u003e Enzyme-labeled antibodies should not be kept at room temperature for a long time. It is best to take them out when used and operate them on the ice box; Enzyme-labeled antibody ( 1× ) Need to be prepared and used now)\u003cbr\u003e④ Preparation of standards ( \u003cstrong\u003e Note: \u003c\/strong\u003e Each experiment needs to use a freshly prepared standard solution): 7uL Of 5ug\/mL Standard is added to 63uL Buffer （ 1× ), formulated into 500ng\/mL Standard, then 64uL Of 500ng\/mL Standard is added to 936uL Buffer （ 1× ), and dilute to 32ng\/mL , and then follow 2 Double ratio dilution as shown in the following figure: \u003cdiv\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231108\/3ecaa9fbfb95470dbb82ee5fc9627285.png\" alt=\"\" width=\"537\" height=\"221\"\u003e\u003c\/div\u003e4 Sample preparation: Return the sample to room temperature and mix the sample before adding the sample; If users need to dilute samples or high-concentration standards in the box, they can use Buffer （ 1× ) for dilution; For cellular samples, it is recommended to press before testing 3000 rpm\/min Rotational speed, centrifugation 5 min , take the supernatant for detection.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e 2. Operation steps: \u003c\/strong\u003e\u003cbr\u003eAll operations are performed at room temperature and repeat assays are recommended for all spiked wells.\u003cbr\u003e1 Returning each component of the kit to room temperature 30min Take out the slats required for the test from the aluminum foil bag that has been equilibrated to room temperature, mark the sequence of the slats with a marker, seal the remaining slats with a sealing film and put them back into the aluminum foil bag, seal them, and place them in 2-8℃ Save. ( \u003cstrong\u003e Note: \u003c\/strong\u003e The slats are easy to fall off during the washing process, so be careful to mark them well.)\u003cbr\u003e2 Sample incubation: add the diluted standard product and the sample to be tested to the enzyme plate (recommended addition sequence: standard well, blank well, sample well, and the standard product is added according to the concentration gradient), 100uL\/ Hole; Enzyme-labeled antibody was then added to each well ( 1× ）， 100uL\/ Hole, seal the plate with a plate sealing film, and then place 37℃ ， 400-600rpm Shaking incubation 1h 。（ \u003cstrong\u003e Note: \u003c\/strong\u003e The loading time is controlled at 10min Within, avoid drift over time. If the plate is not sealed or the plate is not sealed completely during the incubation process, the reaction solution will evaporate, resulting in experimental errors.)\u003cbr\u003e3 Plate washing: After incubation, carefully peel off the plate sealing film, discard the liquid in the wells, and use Buffer （ 1× ) wash plate 3 Times ( 250uL\/ Well), pat dry the residual liquid in the sample well. ( \u003cstrong\u003e Note: \u003c\/strong\u003e If the washing method is hand washing, add Buffer （ 1× ) can be left to stand 1min ； If you use a plate washer to wash the plate, add Buffer （ 1× ) can shake slightly after 5s 。）\u003cbr\u003e4 Color development: adding the pre-arranged color development solution into an enzyme label plate, 100uL\/ The hole is sealed with a sealing film, 37℃ Stand Temperature in the dark 20min 。\u003cbr\u003e5 Termination: adding termination solution, 100µL\/ Holes, can be read after the color is evenly developed ( \u003cstrong\u003e Note: \u003c\/strong\u003e Generally add stop solution 20min Complete readings within).\u003cbr\u003e6 Reading: Put the microplate into the microplate reader and set the wavelength to dual wavelengths 450\/630nm , read the absorbance value ( \u003cstrong\u003e Note: \u003c\/strong\u003e It is recommended that it be set in the microplate reader reading program 5-10s Of shock).\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e 3. Data processing: \u003c\/strong\u003e\u003cbr\u003e1 Calculation of absorbance correction value The absorbance correction value of each standard or sample is: OD450nm-OD630nm- Blank control well absorbance values\u003cbr\u003e2 Taking the standard concentration as the abscissa ( X ), the standard absorbance correction value is the ordinate ( Y ) To draw a standard curve, four parameters are recommended Logistic Mathematical model fitting equation: \u003cdiv style=\"text-align: center;\"\u003eY=((A-D)\/(1+(X\/C)^B))+D\u003c\/div\u003e Substitute the sample absorbance correction value into the formula to calculate the sample BSA Content of. (Note the dilution factor)\u003cbr\u003e3 If the sample to be tested OD The value exceeds the highest point of the standard curve OD Value, the sample needs to be diluted and re-determined.\u003cbr\u003e\u003cstrong\u003e Note: \u003c\/strong\u003e① Per Chinese Pharmacopoeia \u0026lt;9012\u0026gt; 4. Relevant requirements in ligand binding analysis, four parameters are recommended for the fitting equation Logistic Mathematical models. ② Try to use the recommended dual-wavelength correction method, using 630nm Wavelength is corrected, OD450-OD630 Is the corrected OD The value can be directly used for calculation, or it can be corrected for blank space according to the data quality. If there is no dual wavelength microplate reader, read OD450 After the data, the quality of the data should be judged first, and then blank correction should be carried out.\u003cbr\u003e\u003cbr\u003eExample showcase:\u003cbr\u003e(The following standard curves are for reference only, and the standard curves drawn in each experiment shall prevail) \u003cdiv\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231108\/18894eea03e8477486935c9bd2c6b8cf.png\" alt=\"\" width=\"437\" height=\"197\"\u003e\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eBovine Serum Albumin Residue Detection Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e Product background: \u003c\/strong\u003e\u003cbr\u003e Bovine serum albumin ( Bovine serum albumin ， BSA ) is a globulin in bovine serum and has a wide range of applications in biochemical experiments, such as  BSA  It is often used as a stabilizer to restrict or modify enzymes in storage solutions and reaction solutions, and plays a role in “ protect ” Or “ Carrier ” Function, many enzymes are added  BSA  After that, its activity can be greatly improved. Secondly, BSA  Can reduce non-specific binding, often in  Western-blot  With  ELISA  Used as a blocking agent. \u003cbr\u003e Such as partial antibody drugs, vaccine drugs and  CGT  Biological products such as drugs will use fetal bovine serum in the production process, which contains a certain amount of BSA 。 BSA  As an exogenous protein, if it follows the drug product into the human body, it may trigger an immune response or affect the effect of the drug, so in  2020  The third part of the Chinese Pharmacopoeia stipulates that some human vaccines  BSA  Testing requirements: For stock solution testing, it should not be higher than  50ng\/mL For finished product verification, it should not be higher than  50ng\/ Agent. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e Reaction principle: \u003c\/strong\u003e\u003cbr\u003e The kit uses double antibody sandwich enzyme-linked immunosorbent detection technology to determine the sample BSA Trace residue. Coated with capture antibodies  96  Well enzyme label plate, prepare solid phase antibody, then add standard substance and detection sample, and then add horseradish peroxidase ( HRP ) labeled detection antibody to form a solid phase antibody -BSA- The sandwich conjugate of the antibody is detected, washed after the reaction is completed, and then substrate is added for color reaction, and the substrate is subjected to color reaction in  HRP  It changed from colorless to blue under the catalysis of, and changed to the final yellow under the action of the terminating solution. In  450nm  And  630nm The absorbance value was measured at the wavelength ( OD  Values), where  630nm  Is the corrected wavelength. OD  Values and in the sample  BSA  The content was positively correlated. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e Analytical Method Validation Guidance: \u003c\/strong\u003e\u003cbr\u003e1 According to the guidelines of Chinese Pharmacopoeia \u0026lt;9101\u0026gt; 、 \u0026lt;9012\u0026gt; 、 \u0026lt;9401\u0026gt; as well as  ICH\u003cq2\u003e According to the relevant provisions on the validation of analytical methods, it is recommended that all laboratories using this kit verify the following indicators for impurity determination: specificity, accuracy, precision, limit of detection (optional), limit of quantification, linearity, range, and durability. \u003cbr\u003e2 , doing  BSA  When verifying the linearity of raw materials, it is recommended to  BSA  The raw materials are diluted into the linear range of the kit and then measured, and the raw materials are diluted according to a certain proportion to at least  5  The detected concentration is linearly correlated with the dilution factor. \u003cbr\u003e3 Specificity should be verified for other substances present in the process. It is recommended to use high-concentration and low-concentration quality control samples, and add increasing concentrations of relevant interfering substances for specific investigation. The matrix to which no analyte is added should also be measured at the same time. The accuracy of the controls should be within a range and the measured values of the matrix without the addition of analyte should be below the lower limit of quantification. \u003cbr\u003e4 For the accuracy of the sample (spiked recovery rate), please refer to \"In vitro diagnostic reagent analysis performance evaluation (accuracy) - Recycling Experiment) Technical Review Guiding Principles. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e Product performance indicators: \u003c\/strong\u003e\u003cbr\u003e1 Limit of detection: \u0026lt; 0.5ng\/mL\u003cbr\u003e2 Limit of quantification: 0.5ng\/mL\u003cbr\u003e3 Linear range: 0.5-32ng\/mL\u003cbr\u003e4 Accuracy (spiked recovery): 70%-130%\u003cbr\u003e5 Accuracy (measurement deviation): ≤15%\u003cbr\u003e6 , Repeatability (intra-batch difference): ≤10%\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e Specificity: \u003c\/strong\u003e\u003cbr\u003e The type of sample validated has certain limitations, and it is recommended that each user test the cross-reactivity of known substances in their sample matrix in similar experiments. \u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 53.9869%; height: 142px; margin: 0px auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.1332%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003e Antibiotics \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 48.1332%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003e Cross-reactivity rate \u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e Horse serum \u003c\/td\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e Rabbit serum \u003c\/td\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e Collagenase Ⅰ Type \u003c\/td\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e Cysteine \u003c\/td\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e Nuclease \u003c\/td\u003e\n\u003ctd style=\"width: 48.1332%; height: 22px; text-align: center;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px;\"\u003e\n\u003ctd style=\"width: 48.1332%; height: 10px; text-align: center;\"\u003e Trypsin analogs \u003c\/td\u003e\n\u003ctd style=\"width: 48.1332%; height: 10px; text-align: center;\"\u003e ＜ 1%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/q2\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 63.0065%; height: 176px; margin: 0 auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e\u003cstrong\u003e Components \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e\u003cstrong\u003e Specifications \u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e Enzyme plate \u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e8×12 Strip \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e Standard ( 5ug\/mL ） \u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e300uL×1 Tube \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e Enzyme-labeled antibody \u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e40uL×1 Bottle \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003eBuffer （ 20× ） \u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e30mL×1 Bottle \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e Chromogenic liquid A\u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e8mL×1 Bottle \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e Chromogenic liquid B\u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e8mL×1 Bottle \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e Stop liquid \u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px; text-align: center;\"\u003e15mL×1 Bottle \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 31.4869%; text-align: center;\"\u003e Sealing plate \u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; text-align: center;\"\u003e3 Zhang \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. BSA detection is more susceptible to interference and requires high experimental environment. It is necessary to ensure that there is no weighing operation in the environment.\u003cbr\u003e2. Containers and consumables for measuring BSA content should be dedicated to prevent BSA contamination in the laboratory.\u003cbr\u003e3. The pipette is dedicated or uses a gun tip with a filter element.\u003cbr\u003e4. The background value of this kit (i.e. OD0ng\/mL before correction) is generally not greater than 0.2. If the background value is greater than 0.2, the environment, utensils, consumables, etc. need to be checked, and unopened bottled Wahaha water can also be used for water quality inspection.\u003cbr\u003e5. All components in the kit must be restored to room temperature (20-25 ℃) before use.\u003cbr\u003e6. Each component should be thoroughly mixed before use to ensure the uniformity of the reagents. The standard product should be centrifuged briefly for 5s. All the liquid on the tube wall and lid should be concentrated at the bottom of the tube. Immediately after use, all the reagents should be put back to 2-8 ℃.\u003cbr\u003e7. The kit must be used within the validity period, and the corresponding standard curve must be prepared for each test. It is not recommended to use different batches of related reagents in mixed batches.\u003cbr\u003e8. When adding liquid to the microplate, be careful not to touch the bottom of the microplate to prevent damage to the coating layer. Replace the sample loading tank and suction tip in time between different samples and steps to avoid cross-contamination.\u003cbr\u003e9. When patting the slats dry after washing, be careful to prevent the slats from falling off, and be careful not to reuse the sealing film.\u003cbr\u003e10. When the residual concentration of the target to be measured is too high, black flocs may be produced during color development. At this time, the sample needs to be diluted to a certain extent before testing.\u003cbr\u003e11. When reading, pay attention to checking whether the detection wavelength and fitting equation are correct.\u003cbr\u003e12. Only by strictly abiding by the operation methods of the instructions and using all the reagents supporting this kit can the best detection effect be guaranteed.\u003cbr\u003e13. Differences in test results can be caused by a variety of factors, including the operation of the experimenter, the use of the pipette, the plate washing technique, the reaction time or temperature, the storage of the kit, etc.\u003cbr\u003e14. The company is only responsible for the kit itself, and is not responsible for the sample consumption caused by the use of the kit. Users are requested to fully consider the possible usage of samples and reserve sufficient samples before use.\u003cbr\u003e15. This kit is only for in vitro research, not for clinical diagnosis.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplication\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe BSA quantitative detection kit (enzyme-linked immunosorbent assay) produced by our company is a special kit for quantitative detection of BSA residues in intermediates, semi-finished products and finished products of various biological products. The standard product in this kit is a high-concentration standard solution, which meets the individual needs of users; Buffer (20 ×) in the kit realizes the three-in-one combination of sample diluent, anti-detection diluent and lotion in the traditional kit, which is convenient and simple to use. The kit can be directly used for sample analysis, and has good reproducibility and high stability.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe kit should be stored at 2-8 ℃, protected from light, and the shelf life is 12 months. Note that the unused kit after opening should still be stored at 2-8 ℃ in the dark.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41518198915147,"sku":"abs590008-96T","price":714.0,"currency_code":"USD","in_stock":true}]},{"product_id":"protein-a-elisa-kit","title":"Protein A Residue Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e 1. Self-prepared materials: \u003c\/strong\u003e\u003cbr\u003e1 、 10-1000uL Pipettes, single and multi-channel \u003cbr\u003e2 、 100mL And 1000mL Beaker of \u003cbr\u003e3 Deionized water \u003cbr\u003e4 , various types of centrifuge tubes \u003cbr\u003e5 , microplate reader (test 450nm ） \u003cbr\u003e6 Data processing and analysis software \u003cbr\u003e\u003cstrong\u003e 2. Reagent preparation: \u003c\/strong\u003e\u003cbr\u003e1 , Please place this kit at room temperature before use ( 20-25℃ ）。 \u003cbr\u003e2 Wash liquid preparation: 30mL Wash liquid ( 10x ) added to 270mL Deionized water, evenly mixed to obtain 300mL Working washing liquid. If there are crystals in it, please dissolve and mix well before using. \u003cbr\u003e3 Other reagents are used directly without dilution. \u003cbr\u003e\u003cstrong\u003e 3. Sample processing process: \u003c\/strong\u003e\u003cbr\u003e1 If the sample needs to be diluted, use the sample diluent in the kit for dilution, which can be diluted in 96 Orifice plate or EP Conducted in the tube. \u003cbr\u003e2 Ensure that the volume of each well (tube) containing the sample is 100uL 。 \u003cbr\u003e3 Additionally added 100uL Standards and controls into the wells. \u003cbr\u003e4 , adding each well 50uL Acidification solution, mix well by blowing, incubate at room temperature 10min (Make the sample proteinA And antibody acidification isolation). \u003cbr\u003e\u003cstrong\u003e 4. Kit operation steps: \u003c\/strong\u003e\u003cbr\u003e1 Standard dilution:  8  Centrifuge tube markers  1#-8# , take  990uL  Sample dilution was added to  1# And  2# In the tube, take 500uL  Sample dilution was added to 3#-8# In the tube. Take again  10uL  Sample standards are added to  1# In the tube, swirl and mix well before taking  10uL  Then add to 2# Vortex in the tube and mix well. Then take  2# Tube  500uL  Liquid added to  3# Vortex in the tube and mix evenly, then take  3# Tube  500uL  Liquid to 4# Vortex in the tube and mix well, so until  8# Tube. \u003cbr\u003e\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20230707\/fb96890650b64418860723922b09d80e.png\" alt=\"\" width=\"591\" height=\"159\"\u003e\u003cbr\u003e After complete dilution   ， 2#-8# In centrifuge tube  protein A  The concentrations of the standards are  5 、 2.5 、 1.25 、 0.625 、 0.3125 、 0.156 、 0.078ng\/mL 。 Another tube is marked as  0ng\/mL , sample dilution was added. Once the standards and samples are ready, the sample processing procedure is entered. \u003cbr\u003e2 , in the cover  protein A  The antibody is added to the enzyme label plate  100uL\/ Wells of biotin-labeled binding antibody. \u003cbr\u003e3 Re-join  25uL  Acidified standards, samples, controls and blanks into appropriate wells and incubated at room temperature  1h 。 \u003cbr\u003e4 Spin drying, washing  4  Times later, join  100uL\/ Avidin labeling of wells  HRP , room temperature incubation  10min 。 \u003cbr\u003e5 Spin drying, washing  4  Times later, join  100uL\/ Porous  TMB  Chromogenic solution, color development  10min , join  100uL\/ Well stop fluid, then in  450nm  The absorbance value is detected at. \u003cbr\u003e\u003cstrong\u003e V. Data processing and result analysis: \u003c\/strong\u003e\u003cbr\u003e Utilize software such as  Curve Expert  Or  ELISA Calc ) Establish the detection standard curve through the standard concentration gradient, and put  OD The correspondence between the value and the standard concentration is determined by  4  Parametric logistic fitting or cubic polynomial linear fitting into a standard curve, and then the test sample  OD  The value is brought into the corresponding curve equation to calculate the corresponding sample concentration. \u003cbr\u003e\u003cstrong\u003e VI. Analytical method indicators: \u003c\/strong\u003e\u003cbr\u003e1 Precision: at sample concentration greater than  0.3ng\/mL  When, the detection coefficient of variation is less than  10% ； Sample concentration is less than 0.3ng\/mL  When, the detection coefficient of variation is slightly greater than  10% 。 \u003cbr\u003e2 It is recommended that when testing specific samples, the method of adding quality control products should be used to investigate the influence of sample matrix effect on the test results. \u003cbr\u003e3 The sensitivity of this kit is  0.05ng\/mL 。 \u003cbr\u003e4 , hook effect: This kit is a double antibody sandwich one-step method, and the concentration that produces the hook effect is  10ug\/mL 。 \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis kit adopts the technical method of double antibody sandwich labeled with biotin system. The enzyme label plate is pre-coated with a specific monoclonal antibody of protein A. The sample containing protein A is diluted with a sample diluent, and then the acidification solution is mixed to separate protein A from the antibody product. The sample reacts with the protein A capture antibody pre-coated on the enzyme label plate. A biotin-labeled detection antibody is added to form a sandwich complex. Then wash to remove substances that did not participate in the reaction, add avidin-labeled HRP, incubate for a short time, carry out TMB color development, then add stop solution to stop the color development reaction, detect the absorbance value at 450nm, and the OD value is proportional to the sample content.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e This protein A Detection ELISA The kit is used to quantitatively detect unnatural proteins such as recombinant alkali resistance A , can only be used for scientific research and production, and cannot be used for the diagnosis and treatment of human or animal diseases. This kit provides an acidification treatment method of the sample, and the protein A From IgG Isolated from the product, at the concentration of humanized monoclonal antibody as high as 2mg\/mL In the case of protein A The limit of detection is as low as 78pg\/mL Following. \u003cbr\u003e Product Components: \u003cbr\u003e1 、 Protein A Standard （ proteinA  Protein Standard) \u003cbr\u003eRecombinant Alkali-tolerant Protein A in a protein matrix with preservative, 50ug\/mL, 1x0.1mL\u003cbr\u003e2 、 Biotinylated Antibody (Biotin-Labeled Antibody) \u003cbr\u003eBiotinylated anti-protein A monoclonal antibody in a protein matrix with preservative, 1x13mL\u003cbr\u003e3 、 Sample Diluent  (Sample dilution) \u003cbr\u003eTris buffered saline with a protein matrix andpreservative, 1x30mL\u003cbr\u003e4 、 Denaturing Buffer  (Acidification Buffer) \u003cbr\u003eCitrate buffer with detergent and preservative, 1x20mL\u003cbr\u003e5 、 10x Assay Buffer  （ 10x ） \u003cbr\u003e Wash buffer phosphate buffer with preservative, 1x30mL\u003cbr\u003e6 、 Streptavidin labeled HRP  Avidin-labeled  HRP\u003cbr\u003eStreptavidin labeled HRP in a protein matrix withpreservative, 1x13mL\u003cbr\u003e7 、 TMB Substrate  （ TMB  Chromogenic solution) \u003cbr\u003e3,3’,5,5’Tetramethylbenzidine, 1x13mL\u003cbr\u003e8 、 Stop Solution  (Stop Solution) \u003cbr\u003e2M sulphuric acid, 1x13mL\u003cbr\u003e9 、 Monoclonal anti-Protein A Coated plate of 96 wells (coating  proteinA  Antibodies  96  Well plate) \u003cbr\u003e12x8 well strips in a bag with desiccant\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. This kit can only be used for scientific research and production, not for in vitro diagnosis.\u003cbr\u003e2. The terminating solution is 2M sulfuric acid, avoid contact with eyes, skin and clothes. Apart from this, the reagents in this kit will not cause any harm to the human body.\u003cbr\u003e3. High or low PH value, detergent, urea, high salt concentration and organic reagents are all influencing factors of ELISA method. PH has a great influence. The PH value of the sample should be controlled at 7.0-7.4.\u003cbr\u003e4. If you have any questions during use, please contact our technical department.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStored at 2-8 ℃, valid for 6 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41518198947915,"sku":"abs590009-96T","price":714.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/8be814e4d8e946379fb55cca64c0be96.png?v=1789743632"},{"product_id":"ds-rna-quantitative-detection-kit","title":"dsRNA ELISA Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eⅠ. Required instruments and reagents:\u003c\/strong\u003e\u003cbr\u003e1. Microplate reader (including 450nm wavelength, it is recommended to include dual wavelength detection mode, the main and auxiliary wavelengths are 450nm and 650nm respectively).\u003cbr\u003e2. Microplate shaker.\u003cbr\u003e3. RNase-free gun tip and EP tube.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eⅡ. Preparation before the experiment:\u003c\/strong\u003e\u003cbr\u003e1. Equilibrate the reagents used in this experiment to room temperature (18-25 ℃)\u003cbr\u003e2. 20 × concentrated washing liquid is diluted with purified water in a volume ratio of 1:19 to form a washing working liquid.\u003cbr\u003e3. Centrifuge the antibody tube, HRP-SA tube and standard tube at 1000 rpm for 30s before use to avoid residual reagents on the tube wall and cover.\u003cbr\u003e4. 100 × biotinylated detection antibody and 100 × HRP-streptavidin were diluted 100 times with diluent before use and then used.\u003cbr\u003e5. Unmodified, pUTP modified dsRNA standards were diluted to 1, 0.5, 0.25, 0.125, 0.0625, 0.0312, 0.0156, 0 pg\/μL with STE buffer. N1-Me-pUTP modified dsRNA standards were diluted to 2, 1, 0.5, 0.25, 0.125, 0.0625, 0.0312, 0 pg\/μL with STE buffer. 5-OMe-UTP modified dsRNA standards were diluted to 4, 2, 1, 0.5, 0.25, 0.125, 0.0625, 0 pg\/μL with STE buffer.\u003cbr\u003e\u003cbr\u003eDilution methods are recommended as follows:\u003cbr\u003e(1) No modification, pUTP modification\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 81.1138%; height: 239px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 41px; text-align: center;\" rowspan=\"2\"\u003eserial number\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 41px; text-align: center;\" rowspan=\"2\"\u003eFinal Concentration (pg\/μL)\u003c\/td\u003e\n\u003ctd style=\"height: 19px; width: 56.2945%; text-align: center;\" colspan=\"2\"\u003eDilution Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003eSTE buffer\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003eWorking Standard\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e49 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e1 μL 5ng\/μL standard\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e495 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e5 μL of 100pg\/μL solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.5\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL A solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.25\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL B solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eD\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.125\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL C solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eE\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.0625\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL D solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eF\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.0312\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL E solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eG\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.0156\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL of F solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eH\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e\/\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e(2) N1-Me-pUTP Modification\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 81.1138%; height: 239px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 41px; text-align: center;\" rowspan=\"2\"\u003eserial number\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 41px; text-align: center;\" rowspan=\"2\"\u003eFinal Concentration (pg\/μL)\u003c\/td\u003e\n\u003ctd style=\"height: 19px; width: 56.2945%; text-align: center;\" colspan=\"2\"\u003eDilution Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003eSTE buffer\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003eWorking Standard\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e49 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e1 μL 5ng\/μL standard\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e490 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e10 μL of 100pg\/μL solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL A solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.5\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL B solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eD\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.25\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL C solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eE\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.125\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL D solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eF\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.0625\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL E solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eG\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.0312\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL of F solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eH\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e\/\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e(3) 5-OMe-UTP Modification\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 81.1138%; height: 239px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 41px; text-align: center;\" rowspan=\"2\"\u003eserial number\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 41px; text-align: center;\" rowspan=\"2\"\u003eFinal Concentration (pg\/μL)\u003c\/td\u003e\n\u003ctd style=\"height: 19px; width: 56.2945%; text-align: center;\" colspan=\"2\"\u003eDilution Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003eSTE buffer\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003eWorking Standard\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e49 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e1 μL 5ng\/μL standard\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e480 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e20 μL of 100pg\/μL solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL A solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL B solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eD\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.5\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL C solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eE\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.25\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL D solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eF\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.125\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL E solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eG\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0.0625\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e250 μL of F solution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.6252%; height: 22px; text-align: center;\"\u003eH\u003c\/td\u003e\n\u003ctd style=\"width: 23.3219%; height: 22px; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 22.9722%; height: 22px; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 33.3223%; height: 22px; text-align: center;\"\u003e\/\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cstrong\u003e\u003cbr\u003eⅢ. Experimental steps:\u003c\/strong\u003e\u003cbr\u003e1. Take out the required slats from the aluminum foil bag after room temperature equilibrium, cover the remaining slats with a sealing film, seal them with a ziplock bag and put them back at 2 ~ 8 ℃.\u003cbr\u003e2. Set standard wells and sample wells, add 100μL of standard substances of different concentrations to each standard well, and add 100μL of sample to be tested to the sample well.\u003cbr\u003eNote: When the dsRNA content in the sample to be tested cannot be determined, the STE buffer should be used to make multiple dilution factors for detection, so as to avoid the content being too high and the valid value cannot be read.\u003cbr\u003e3. Seal the reaction holes with a plate sealing membrane, and vibrate the plate (500rpm) for 60 minutes at room temperature.\u003cbr\u003e4. Discard the liquid, pat dry on absorbent paper, fill each well with washing liquid (250μL), let stand for 30s, throw away the washing liquid, pat dry on absorbent paper, and repeat washing 4 times.\u003cbr\u003e5. Add 100μL of biotinylated detection antibody at working concentration to each well of the standard well and sample well, seal the reaction well with a plate sealing membrane, and vibrate the plate (500rpm) at room temperature for 60 minutes.\u003cbr\u003e6. Discard the liquid, pat dry on absorbent paper, fill each well with washing liquid (250μL), let it stand for 30s, throw away the washing liquid, pat dry on absorbent paper, and repeat washing 4 times.\u003cbr\u003e7. Add 100μL of HRP-streptavidin with working concentration to each well of the standard well and sample well, seal the reaction well with a plate sealing membrane, and vibrate the plate (500rpm) at room temperature for 30 minutes.\u003cbr\u003e8. Discard the liquid, pat dry on absorbent paper, fill each well with washing liquid (250μL), let stand for 30s, throw away the washing liquid, pat dry on absorbent paper, and repeat washing 4 times.\u003cbr\u003e9. Add 100μL of one-component substrate chromogenic solution to each well, seal the reaction well with a sealing film, and let it stand at room temperature to protect from light for 30 minutes.\u003cbr\u003e10. Add 50μL of stop solution to each well, detect immediately, and set the wavelength of the microplate reader at 450nm (dual wavelength 450nm\/650nm is recommended).\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eⅣ. Interpretation of test results:\u003c\/strong\u003e\u003cbr\u003e1. Take the average light absorption value of the standard product, the blank control, and the sample, and subtract the average light absorption value of the blank control to obtain the light absorption calibration value of the standard product and the sample. The standard curve is drawn with the standard concentration as the abscissa and the calibrated standard light absorption value as the ordinate. (The light absorption value in the dual-wavelength detection mode is 450 nm minus 650 nm)\u003cbr\u003e2. It is recommended to use professional curve making software, such as curve expert 1.3 or ELISA Calc (please use Logistic five-parameter or four-parameter fitting curve calculation method to draw standard curves), etc.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eⅤ. Detection performance\u003c\/strong\u003e\u003cbr\u003e1. Detection limit:\u003cbr\u003eNo modification, pUTP modification, N1-Me-pUTP modified dsRNA detection limit ≤ 0. 001 pg\/μL, 5-OMe-UTP modified dsRNA detection limit ≤ 0. 01 pg\/μL.\u003cbr\u003e2. Limit of quantification:\u003cbr\u003eNo modification, pUTP-modified dsRNA limit of quantification was 0.0156 pg\/μL, N1-Me-pUTP-modified dsRNA limit of quantification was 0.0312 pg\/μL, and 5-OMe-UTP-modified dsRNA limit of quantification was 0.0625 pg\/μL.\u003cbr\u003e3. Precision:\u003cbr\u003eIntra-plate coefficient of variation ≤ 10%, inter-plate coefficient of variation ≤ 10%\u003cbr\u003e4. Recovery rate:\u003cbr\u003e80%~120%\u003cbr\u003e5. Linear range:\u003cbr\u003eThe linear range of unmodified, pUTP-modified dsRNA detection was 0.0156-0.5 pg\/μL, the linear range of N1-Me-pUTP-modified dsRNA detection was 0.0312-1 pg\/μL, and the linear range of 5-OMe-UTP-modified dsRNA detection was 0.0625-1 pg\/μL.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis kit adopts the principle of double-antibody sandwich method and is coupled with biotin-streptavidin system to quantitatively detect the content of double-stranded RNA (dsRNA) in samples. The length of the detected dsRNA is 60 bp or more, and the detected dsRNA is independent of its nucleic acid sequence. The microwells of the plate were coated with anti-dsRNA antibody, incubated and washed, and then incubated with biotinylated detection antibody to form antibody-antigen-antibody complex. After washing again, horseradish peroxidase (HRP) labeled streptavidin (SA) was added. After thorough washing, the substrate TMB was added to develop color, and TMB was converted to blue under the catalysis of peroxidase and to final yellow by acid termination.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eDouble-stranded RNA ELISA test kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003ctable style=\"border-collapse: collapse; width: 65.8596%; height: 330px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003eSerial number\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eReaction plate\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e8 × 12 wells\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eBiotinylated detection antibody (100 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e120 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eHRP-Streptavidin (100 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e120 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eDiluent\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e30mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e5\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eChromogenic liquid\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e12mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eStop liquid\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e6mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e7\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eConcentrated wash (20 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e40mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e8\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003edsRNA standard (no modification, 5ng\/μL)\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e15 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e9\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003edsRNA standard (pUTP modified, 5ng\/μL)\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e15 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e10\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003edsRNA standard (N1-Me-pUTP modified, 5ng\/μL)\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e15 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e11\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003edsRNA standard (5-OMe-UTP modification, 5ng\/μL)\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e15 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e12\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eSTE buffer\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 11.6605%; height: 22px; text-align: center;\"\u003e13\u003c\/td\u003e\n\u003ctd style=\"width: 64.4775%; height: 22px; text-align: center;\"\u003eSealing film\u003c\/td\u003e\n\u003ctd style=\"width: 18.7709%; height: 22px; text-align: center;\"\u003e4 sheets\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. The color development temperature and time are crucial to the experimental results and should be accurately grasped.  \u003cbr\u003e2. During the washing process, the washing liquid should be soaked in the reaction plate for 30 seconds and then dried to fully wash the non-specifically adsorbed components.  \u003cbr\u003e3. All reagents should be thoroughly shaken before use. When adding samples, the added sample should be added to the bottom of the well of the enzyme label plate to avoid adding it to the upper part of the well wall. When adding samples, be careful not to spill or produce bubbles.  \u003cbr\u003e4. If crystals are found in the concentrated washing liquid, they can be incubated in a 37 °C water bath, and then mixed and diluted to the working concentration after the crystals are completely dissolved.  \u003cbr\u003e5. Avoid introducing sodium azide (NaN) into the sample\u003csub\u003e3\u003c\/sub\u003e), sodium azide will destroy horseradish peroxidase activity and make the detection value low.  \u003cbr\u003e6. RNase contamination should be strictly avoided during the experimental operation.  \u003cbr\u003e7. Do not use a shaker instead of a vibrating plate machine. If there is no vibrating plate machine, room temperature incubation can be used, but standing incubation will cause the detection sensitivity to be about doubled.  \u003cbr\u003eIt is recommended that unmodified, pUTP modified standards be diluted from 2pg\/μL, N1-Me-pUTP modified standards be diluted from 4pg\/μL, 5-OMe-UTP modified standards be diluted from 8pg\/μL, and the HRP-SA incubation duration be adjusted from 30min to 60min.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. The kit is stored at 2 ~ 8 °C, avoid direct strong light, and is valid for 12 months.\u003cbr\u003e2. After the coating strip is unpacked and used, the remaining coating strip should be sealed, stored at 2 ~ 8 °C, and used within the validity period.\u003cbr\u003e3. Other components of the kit should be put back to 2 ~ 8 ℃ in time after use and used within the validity period.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41518199308363,"sku":"abs590014-96T","price":1250.0,"currency_code":"USD","in_stock":true}]},{"product_id":"ua-myco-nest-pcr-mycoplasma-detection-kit","title":"UA-Myco Nest PCR Mycoplasma Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eNested PCR Mycoplasma Detection Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eIce pack transportation. Store at -20°C away from light, valid for 18 months.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eThe UA-Myco nested PCR Mycoplasma Detection Kit is designed for the qualitative detection of Mycoplasma contamination in cell cultures. Over 95% of Mycoplasma contaminations in cell cultures are caused by Mycoplasma fermentans, M.orale, M.pirum, M.hyorhinis, M. hominis, M.salivariu, M.arginine, and Acholeplasma laidlawii. This kit amplifies conserved gene fragments of these eight and other common Mycoplasma species in cell culture media through nested PCR, thereby determining the presence of Mycoplasma contamination. The kit features rapid detection, high sensitivity, and high specificity. Each reaction can detect as few as 10 Mycoplasma gene copies, with particularly improved sensitivity for Acholeplasma laidlawii, and shows no cross-reactivity with bacteria or cultured cells.\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003eThe Nested PCR Mycoplasma Detection Kit includes 2 tubes of ready-to-use primers and 1 tube of reaction internal control, with the following specifications:\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 674.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 150.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eSpecification\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 523.009px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eComponents\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e25T\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003ePrimer\u003c\/font\u003e \u003cfont\u003eP1 (50 μL), Primer P2 (50 μL), PCR reaction internal control (50 μL)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e50T\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003ePrimer\u003c\/font\u003e \u003cfont\u003eP1 (100 μL), Primer P2 (100 μL), PCR reaction internal control (100 μL)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1. \u003cfont\u003ePreparation of Cell Culture Medium for Testing\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1) \u003cfont\u003eTake 1 mL of cell culture medium from a cell culture in the logarithmic growth phase.\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2) \u003cfont\u003eCentrifuge the cell culture medium at 200g for 5 minutes at room temperature. Transfer approximately 800 μL of the supernatant to a new centrifuge tube, taking care not to transfer the pelleted cell debris.\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e3) \u003cfont\u003eUse the prepared cell culture medium as the template for nested PCR to detect mycoplasma.\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2. \u003cfont\u003eNested PCR Reaction\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1) \u003cfont\u003eFirst PCR Reaction: Using a standard 2X PCR Master Mix as an example:\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003ePCR Reaction Setup (Recommended to prepare on ice)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 492.41px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 253.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 238.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eVolume\u003c\/font\u003e \u003cfont\u003e( μL）\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e2xPCR Mix   \u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e25\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:5.0000pt\"\u003e\u003cfont\u003est \u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003ePCR Primer P1\u003c\/font\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eTemplate (Cell Culture Medium)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003ePCR\u003c\/font\u003e \u003cfont\u003eInternal Control\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eAdd sterile ultrapure water to\u003c\/font\u003e   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e50\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003ePCR Reaction Conditions: 30 cycles, see the table below for conditions\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 512.41px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 128.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eStep\u003c\/font\u003e  \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 140.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eTemperature\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 110.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003eTime\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 133.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eCycles\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eInitial Denaturation\u003c\/font\u003e   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e94℃\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e5min\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eDenaturation\u003c\/font\u003e   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e94℃\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e30s\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e30\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eAnnealing\u003c\/font\u003e   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e55℃\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e30s\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e30\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eExtension\u003c\/font\u003e   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e72℃\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e40s\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e30\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003eFinal Extension\u003c\/font\u003e   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e\u003cfont\u003e72℃\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:11.0000pt\"\u003e \u003cfont\u003e5min\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2)  \u003cfont\u003eTake 1 μL from the first PCR reaction as the template for the second PCR and perform the second PCR using primer P2. The setup for the second PCR reaction (recommended to prepare on ice) is shown in the table below. The PCR reaction conditions are the same as the first PCR reaction.\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 504.41px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 233.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 270.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eVolume\u003c\/font\u003e \u003cfont\u003e( μL）\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e2xPCR  Mix   \u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e25\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e2nd PCR Primer P2\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eTemplate (1st\u003c\/font\u003e  \u003cfont\u003ePCR Product)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eAdd sterile ultrapure water to\u003c\/font\u003e   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e50\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e3) Prepare a 2% agarose gel. Take 10 µL of the second PCR product for DNA electrophoresis.\u003c\/font\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e3. Results\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eThe PCR internal control yields a single DNA band at 515 bp. Common mycoplasma contamination produces a single DNA band between 236 bp and 365 bp, while Acholeplasma laidlawii contamination produces two DNA bands at 426 bp and 219 bp.\u003c\/font\u003e\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003eIt is recommended to use hot-start Taq enzyme series for PCR reagents. The 2X PCR master mix should not contain DNA gel electrophoresis dye.  It is advised to assemble the PCR reaction in a clean bench to avoid cross-contamination and aerosol contamination. Use nuclease-free pipette tips and reaction tubes, and filtered tips are recommended.  Unless strictly validated, it is not recommended to arbitrarily alter the reaction reagent volumes.  Store the reaction reagents according to the instructions to ensure reagent stability.  This product is for research use only.\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"25T","offer_id":41518217756747,"sku":"UA079017-25T","price":30.0,"currency_code":"USD","in_stock":true},{"title":"50T","offer_id":41518217789515,"sku":"UA079017-50T","price":50.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_d356060f-ac30-4795-86e5-f84f4dd24b0d.png?v=1789113624"},{"product_id":"startscript-1st-strand-cdna-synthesis-kit-with-dnase-i","title":"StartScript® 1st Strand cDNA Synthesis Kit, with DNase I","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e[1] Roth, M J ,  N. Tanese , and  S. P. Goff . \"Purification and characterization of murine retroviral reverse transcriptase expressed in Escherichia coli. \" Journal of Biological Chemistry 260.16(1985):9326.\u003cbr\u003e[2] Kotewicz, Michael L , et al. \"Isolation of cloned Moloney murine leukemia virus reverse transcriptase lacking ribonuclease H activity.\" Nucleic Acids Research 16.1(1988):265-277.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 863.604px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 351.863px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 260px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eUA070077-10 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 251px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eUA070077-50 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eDNase I, RNase-free (2 U\/μL)  \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10X Reaction Buffer with MgCl2 for DNase I   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e50 mM EDTA   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eM-MLV Reverse Transcriptase (200 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e5\u003c\/font\u003e0 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e5× Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e40 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNase Inhibitor (40 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e5 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e25 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003edNTP Mixture (10 mM each)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e5\u003c\/font\u003e0 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eOligo(dT)18 Primer (50 μM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e5\u003c\/font\u003e0 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\n\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRandom Hexamer Primer\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e(50 μM)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e20 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNase Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003eI. Removal of genomic DNA from RNA preparations\u003cbr\u003e1. The reaction system is formulated as follows:\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 712.604px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 432.863px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 279.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNA\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10X Reaction Buffer with MgCl2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eDNase I, RNase-free (2 U\/μL)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNase Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eTo 10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e2. Incubate at 37 °C for 10 min.\u003cbr\u003e3. Add 1 μL 50 mM EDTA and incubate at 65 °C for 10 min. RNA hydrolyzes during heating with divalent cations in the absence of a chelating agent. \u003cbr\u003e4. Use the prepared RNA as a template for reverse transcriptase.\u003cbr\u003eII. First Strand cDNA Synthesis\u003cbr\u003e1. Set up the reverse transcription reaction according to the following table\u003cimg class=\"img-fluid\" 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iIAcKkWrqk8fEBDh+4BFf1LcEltz0NN0On337h2PdXr16hU25E4mE6azLPv111950BOlgdvB9i9fvqQQnA2FfbjQAft2ufM3yzLu2+0PXKPVJLH4xHPGOh1m0UepcAhFz+4MjwEIcyjKejF8jXvkrUPZbe8WATeXX5ZlTVvz9R+bjiaOqU9Ugf5nJuZQPQQcI19f0TuVaUT4ToX8kdq183ZHalLNHAuBZOITwoB9lsjzvIvDYcH2tkrmOgdrSbiS+yosNHeZrGXJJUqS85VliRynzHK5vLi4WCwWFIJEtHWWubq6CqvwKhNH04ctDiuRjEP1E3ZONIjwncpAInynQv5I7YrwHQnoUzSTTHyyawYIZJ7nlh4x\/8DEVo9YLBZnZ2dFUbiGwEEtK0UB8LZwQs5uikdJTOCRHeJ40O+++852050ZSh2qWscCjKurK5asSVjCV5ZlR\/rUKNDzS8k41Mlxns1mVawuz\/OqSydXO3kFRPhSNrF9Q8Qwk3KHR9a3NOITGJh9swkzgp2E+QcaeSvhW1Gr0FkWi8XFxcXr169Xd8zwqjvokxo2IVj39\/fT6ZQhMEr4trYepZtUg4nj6MPmBpdIw6FOCDseS9yktdNHhM8BcsyfInzHRFttCYE2EUgjPrl5JgIEZhYeGGPn2MBgomfJ2HemWZaROELsb7\/9hrNnSOAQq7799tsvvvgi5E9rEphjMo\/kzKka1rIEC+1a5cntgAA0ZCaFl2VZ3\/rNzQ3O1Kk5dGe5XLoZx+70sZoPK52GQx0fc84shM9CoTL1hM9ODc7nc+svdHawFjsvHrainCgCInxRWJQpBAaAQBrxqYrwgV3hjg92glu841X2kj1LBmlEIJQBo0IaQmbrP1gaTOuvv\/6aTqfufa5dVwde6HSAqmG0syEqjE+W21GszaRiXNXHYhydq+7c3t5yZz1aQbs2DYWPoA8VG2IiDYc6MvLWiZo0vR\/hs27LL0s0aU5lLAIifBYNpYXAkBBIIz7VEz5Mp9nbvbOQuwROM5\/P3caL+XwOAmTL2zIrNfI8d5MKaGs+n7sJwnCSLxr2rDKrTfSOEVpuR3Z7f39\/dXWFCbkmrd\/d3YGe2n5VpY+gj7POsH6m4VDHxxwuTB+pV2A\/wmdd1abr29JVh4AInwNEP4XAYBBIIz6BnYTRAuwknJZz5rHkhhNdoG52Uo1BwpXnKyRsdA0JH6jY5kb5+J+VDH3qCR9ey7palvBR86+++ury8pKEr751K8H2qypNwteRPs40g\/uZhkOdCnbQvpWLuAcbp89+hM9OoodO6prQzyoERPiqkFG+EOg7AsnEp627dC2DcVZxl\/jTkhssXzs7O1us\/6bTKd\/JotjLly+vr69RzL3SLYri+fPnZGBcCWcZqo1GVj2nA8klyli6hhyETDt9WN96URRQm3wR\/SIILr9rfWzfh5hOxqFOCz4WHthhbPXZj\/DR77Iscw9OVrjS9QiI8NXjo6tCoL8IJBOfQJjsAm23WA0MJjpzYMmNPUvGyeQMnCuPKgwh7iqEkB1yKDiG6mqxmCNYKMZu2qWBrGJJ4dbWr6+vudzQ6lCV7lof9mKgiWQcqs\/42xEe6jmbzeAg9N\/7+\/ubm5voh9DD6sqpR0CErx4fXRUC\/UUgsfgEkodbElmRfbK3m21pFZCbzY0sy+y8AmIGLtlXw8ghfVwsFvY7dnmeo7CtzsI2k5+fiH6WxZ5sQa342uvLL7+kzpZQctrPNlTV+t\/+9jdMPRIEe9COTeOUHbbYkT62IzTQgBKJOVTfkOcorZmlo9dMJpM3b97gFKj5fG7dAcPY3iL61tPe6rO5A\/RWv7YUc88HvO12eoiTVZ7jmPduXnWP3cxvnjhcQlVb3Ulmiy02QVE1h1Cx3QQSik\/uleV+NrXzZBAY\/fBylXCEoqFznarejSpfDnVyc1fduvkgBA3tHPbJdR6QAqMgfFyvzU9z2YcD9\/KoC+OtmgDPC\/eTg6NwqmCP1g+XUNMoKVRNmQMvtdWExQExOOTWB6rat+qKT4cTPje9BxPbO0a90Ucy0upBSOaqHOq0pqyhcW4SPYykp9V8KK2nT\/jcQUlu8Q3s1NEhThBeNQFQFMXd3d0hEetwCSccplS+RR3s\/aIK9habO7koxSc7Vb8rv8eTnn32swaNLrCzBZCezWaa2wthGWiOHOq0hsMNvMqR+ZYM6xPsPqrTqj2g1tMnfJxdI7Wy2+tgKkwOhfmtGNKyEAq001rRAixZlThcQpXkI+Rb5VtsziHpHgpbbKgnohSfemIIqZEGAnKoNOyoXlQhkDjhCxnAqsPhAwSKnZ+fv3\/\/nqtE8TzB1c3YzQe8otQQU0qugJ2BcDsBUXI6nb579w7fiWjeIhSukYCjlnjkFKcx+JBkuxDNxIjBJcxhYEakatVj2H2wOurAhbpO+RcvXvCNdiiEhfHxW\/tsR7VpIxTmjIv7WeUDw81XfBqu7aR5DxGQQ\/XQKFKpRQTAGbIWJfZKlFuLA\/oVEj7wDLAiyzDsRBTf3VTRiBX\/gGQUIKOKlreZSIP0NG9xqwQUAMsEB4p2IZpJI7IV+z0wfKKWZZCIdt\/qAMZsQYZWbAIFQgxRwOFDy1rErCiu1wzN7TQf7k\/Fp+HaTpr3EAE5VA+NIpVaRCBxwuc+yV1P+DBRZEkD0+QcG7zMWewwBvkHfvIcJ75H5rQTClCyK8D8rS2yZL0EthsVGM20w4utgFeRxdoy4edq2X1WR3mSM5vPdBWGLOB6Wo8krvLra07hNH4qPqVhR\/WiJwjIoXpiCKnREQLjInygDiFrsWwmSi8cFwmNQYqDS7a8FciKNjOathJYyyaitSwlsgVCThbqaYUzbYXw9XQ4Z1bVfVuduuV5bvOZbiKEhSkNIzicLEQXRPhoSiWEgBCoR0CErx4fXR06AokTvpA2bd2lG1IKHMQ+mUw4WxZanXNXuFQUBc5xIi9xdaOt2MJOYNjiVgm2AL9e69TYqRXoEAXQyWH3nQ78yURNl6NCWJGWZY4VBVXxpj6kpyGYA81RfBqo4aR2PxGQQ\/XTLtKqLQQSJ3yWDQAyu1wPOdiLQCZkq9i0JTqr\/NvbW2sDJ9ZOLDkyhFqQnOd5URS\/\/\/47D\/dkfnjik2uRJasksAD1jHYhmskqEALKe3l5CVIVfpa2qvusDoE8VIe6WeWbCKFAfoeJOSHhs5fYo5QSik8pWVN9OTkCcqiTm0AKdIpA4oSvLEselGRx5IZTt+cUFAH7SV+9ejWZTABQnuegI\/jJTa9Wpi3At8Z8Deoa4p7fr7\/++vPPP9+vRWxTrZLw2WefQVsqYzVkF6KZ6BfRyLLs559\/tmjYjiNt5bBFK8FusGX3v\/322y+++AJ6uvNzIIQS3DlRL1++xH7qyWTy9OnTLMtevHiBnJU0zOrpsyyhmUaSU+N3R0YgfN7j7nK6SY1KLAwf4VZ37klyPl6Wpe07n2PZRNVJBiww5oQIX7vWZ5wNxyEbgoOEw5gFlGgRgc1tpEWJfRPlPrzcN\/XS1gd0rcbbu+s+COhJmu6uU06y4pMDhDSIzzN45ON3f8LyXec4wrcKgZjX\/+yzz548ebJYLLYqYCVgVPMDT+yv62DVV6PhjwkvctgKZn0BOVQ9Pjtd5Y7J+igQ\/ebDTg2pcHME0id8VfsVmmOkknsjUO\/qe4vdWhFxMfnApvgUjgS7RIFXOz1Kh61sTez3EGIJHyfwOLZxxuh3333nWODV1VV4FAFmXCwb3qrzqArIofYzN9fqsLob6lUvW4qiuL6+Zi0lukZgFIQPz8EXFxdNnqe7Rnw88u2rJcan43SfHxc8TnOnakXxySGPB4zwVSk4U5jvqnf9c7\/nnyaE7\/7+fjqd8gvk7qR59GuxWFxcXLx+\/Xp10z+yP3YNbFvy5VA7IYnnBze7DAncVBf9yVa4YiHtVzHs78kTYyF8JwdaCgiB1hFQfHKQ4hkjZDNgWufn51Un0FAOY9h0On379i3Wrb5+\/fr8\/Nyun0N5hitLJZmJiTT8nK\/\/NnfbLPv0009tmHRVqAwSlvChI3aKjtwOfYcmzLSiHt4lP65FJjW0BZSWQzUZA3ySDx2N1fk+Fzn1jzp26FICE3b60ImFa9Ct7CQ3qythERDhs2goLQSGhIDik7NWFeHDCybwJMQeewINhYTHydjC2GlEssVZZBvMuD0fmVWfnKyvQn2QsFEtDGmW24GtYvOTe6VrV\/WxmGtIP+VQW8cAR3h9ScfM7ICPVqwpUEX4bBV+tCEqXJlEQISPUCghBAaGgOKTM1g94QuP0nHVQa3sdJ0NKvZjlsjn1EK2PnrHvcaCcCuBaSaiVaJa8WugblrFEj7y2vv7e0f4cBS1VUmTfA7n1f4eOVSISZhTPyGH8o7w2VnqUCByot\/TwFMWx6oVa33HpqvkKx8j\/OGuJSyEgBAYHALyXGcyECnL2FDAMjmSLVcXPxHMVvdE8CpXGD+xzXY6nRZFYYXYaMR8K4FpJqJVWBcJGyzD3SeW8PFTlF999dXl5SU3bYAIWnoavp52jY7zpxyqud2dp7iKjoG5n64wftpnElugaoYPoxp+2sSPrMzRpjc3gdH2Xx0XAsNFQPEptN3WXbokW2Fd5lCIK8yfloSxVjSqsQrZ2Hw+Z2a0CgUi4dqy8Q\/b0cLJPPfNS\/uq2soMmbFremw\/5VB7WBwrBDgJBwlgY9yKkee5KxA2lOc5y9urdsA7YgfX0NOLhas+LcJXj4+uCoH+IqD4FNqGrzW5Jd8tWQPZci9GIacoCnecDJkZCtjYQ1IIJnd7e+uaXiwW+MYYz9EhdaPYaBXXKdZCPupyKaFdnMeKVk8Xem2ZcEUgr44zIYdq0e78FAtHe43w8JmEhWezGUY7neX+\/v7m5maxWDx79oxuzvJK1CAgwlcDji4JgV4joPhUZR6QPNzdyI24CA\/54fwWJwxWBewr3c1dMsvsLAXCj2vCZb57946nv\/zwww+U88knnyDtTpexqqJr3MBrJ+34Nu3LL7+kTDs7wve8Vh9yXJtpNwtXgTmefDlUu7amG3JwYuzBj6y7Wc9yOtAFJpPJmzdv4FDOcZwbOgn6SQQ2twv+VkIICIGhIKD41LWlmkxOdK2D5B8NATnU0aBu3lDVYYB8qoEouWoTSEX4mqCkMkKgjwgoPnVtFUWRrhHulXw5VK\/MYZe9horxlTEu6cssIURhjghfiIlyhMAwEFB86tROfHO6ukvyfWinLUr4aRGQQ50W\/7B1PHFVeR\/f9toFD6EQ5RABET5CoYQQGBgCik8DM5jU7TcCcqh+20faHYrAhvDpPyEgBISAEBACQkAICIHEETiUN6q+EBACR0dAExJHh1wNpoyAHCpl66pv67NkHrisoBACQmBwCMhzB2cyKdxnBORQfbaOdDscgc3M5eGCJEEICIEjI6D4dGTA1VzaCMih0raveifCpzEgBIaKgOLTUC0nvXuJgByql2aRUq0hIMLXGpQSJASOjIDi05EBV3NpIyCHStu+6p0In8aAEBgqAmOOT\/Yjee5EMh4+y8R+Bo4e1kmZ9htgm9vo+r\/pdPr3v\/+9KIqaRu0BU7YYZYbHvtli9WlqWF9MV0MExuxQIRpt5VSN9q3yNZK3QrRrARG+XRFTeSHQFwTGGZ8QPyzJw5GdOK8TQQJHxB4SMIqimE6njrdR+G+\/\/fb8+XNcta2AI75\/\/\/78\/Lzqa7H4lmz4qdjZbMYTRWfrv\/3GmdVnPwmjrTVOh+rU3FWjfWuj9DWew7u1igpsRUCEbytEKiAEeorAOOPTbDabTCaOiuV5zszDz0NbLBZnZ2euCQwCxKHffvvt5ubG5jAs3d7eLhYLsFJmhgMoz3PSu7IsUZ4c0R0bFVZXThcIjNOhukDSyXSj3V2t+nm4I1dJHm2+CN9oTa+ODx6BEcYnxIDwjSd4GPIPjxOrKUNyLzdKQuFVM2r1pM2FQPfaq0YBp49+tojACB2qRfRqRLnRXlPSXgp9zV5Veg8ERPj2AE1VhEAvEBhhfMLSvZCNITZgzoxxAkvi3Kte3PIwHQiahRySSFa3Nl4xMBR7+vTpZDKxU3dVhC8qhzLDEEjOulgsLi4ultnvnaUAAB35SURBVMslC5dliatZlr1+\/Xr1vjjLMnTB5U+n0xcvXmRZ9vPPP0\/Wf3\/++SfKz2Yz9tdOLtqFg6G0xWJh1Ug7PUKHOo5Bw9Fu27WLGebzOVdr1HuQlaB0QwRE+BoCpWJCoHcIjDA+VRE+UBmECsaJMFGWZZ7njCgrrgPuiJLgfOHqPVsFzK8J4YNKITfFMIqGQPSO6rkBByX55no2m9n+In8+n7teW15o0+jCbDZDIlrLdtMpk+TPETrUcewYHe1sWoSPUHSdEOHrGmHJFwJdITDC+FRP+NwMH0mM5XBMMwHzzOdzciZLuSgExdxPzr1FuVHN3oswBC6Xy4uLi\/\/+97\/T9V84teaaxmycZXhOQ1s+mkbmJgas\/5vNZrZkVwO3r3JH6FDHMUU42m27InwWjU7TInydwivhQqBDBEYYn0BH+PqV4IL9uDV85C52so3vjMjwIIT8jwVcPn5Spmv6cMLH6cayLG0IZEOuafzM8zya74igLcM0u8wmyrLkVZs5kvQIHeo4lhXhOw7OW1sR4dsKkQoIgZ4iMM741HyXruUuXJ2GabxwZq4oirOzs8Vi4WgQJ9IwCKxM5LgCHCuWZTKTCRcCndjoJwDDMtPp1BE7y9hs+Wg6qrktSW1HkhinQx3BuG60uxbt44193BrzUHQQtfVThK8tJCVHCBwbgXHGJ7tcD4i7dXWMEzbx7Nkz95LUyeHrV9aCcBSrookhceQgcHKYj4QNcvwsC\/dSRHf4OoGUgHwuFmQxJiwLdGnLnlfl8U2Z6XRKaU7ttH+O06GOYFOO1WhbXI1Kl7y\/v7+5ubEDOFpRmbsiIMK3K2IqLwT6gsCY4xO3zWZZFi65w31tMpkg8dtvv2Gn6uZ+91gFAQaZ9jVxnuf2py2GXbr4sDPevVIm6RrGR5S0kW+hFnkkOR\/ynSgIRPxzzXHmMssyrsBDmSdPnmAz76tXrwDFZDJhGl2wXZtOp2\/fviVoFoG+jPiO9RizQ3UErR20drTb5rhPfDKZvHnzZvW8kWXZr7\/+isRqMI\/z8cNC1FZ6c\/doS5zktIsAPYF3+aq7cPTtTLvKSFrfEFB8amiR5XJ5dXXFD51snTmo+fBy8xbPz8+jC\/saSgiLbVU7rKKcnRCQQ+0EV1uFLy8vox85b0u+5BABET5C0dOEZXLYohjlfLZYT3sitdpGQPGpIaJuss1+ZqVKglvJV1Usmo9ps9anJUT4omi3mCmHahHMhqI0qhsC1UoxEb5WYOxQiGNyds1Nh61K9BAQUHxqbiU7WR59YRqKwndS3Mq\/sFiYw4\/bhZf2zsHDHu7XrVPJvbVKrKIc6vgGBeHTkD4O8iJ8x8F5\/1Yc4XPr0\/eXq5rDR0Dxafg2VA96hIAcqkfGkCodICDC1wGorYp0hA8zfK9fv8baapyz1MV5SlwUb1fEt9ozCTsUAcWnQxFUfSFgEJBDGTCUTBABEb6+G9USPrzWwQspzIRj35P9FpfNR12UsXL4yonrJ2yt+XxeFMXz58+xyP3q6mqPt1p9hzUJ\/RSfkjCjOtEXBORQfbGE9OgGARG+bnBtTyqI2sZOWcblR+RqaIo\/meAHILBVkPlIUKD9mgM3FaLR6O6Q9nomSYcioPh0KIKqLwQMAnIoA4aSCSIgwtd3o9qZOasrCRwy+ZOJKsIX3X5oa0EgF4lrOa2FvVdpxademUPKDB0BOdTQLSj96xEQ4avH5\/RXuyB8k8mEk3noYUj4kK9NwacfAdUaKD5VY6MrQmBnBORQO0OmCoNCQISv7+aqJ3ycfiNjY6Jqhg\/HA\/Cj56vy4XlKRVFcXl5CwtnZmb6K2c9RovjUT7tIq4EiIIcaqOGkdkMERPgaAnWaYlUfDwMLhPG6OE\/JyienPA0EarUagWHFJy4ScIehVffv4Yp1AQx4PqvUV6y\/WvUcVV+r6qpVEqtsw5yqun3ObxelPvcUug3LoXbFE98D5yrw5tXrh8FiseBB1fy2g3uDFLblvoUeFghztqrBo9i2tu6EU3LyZ36I8DnT66cQGAwCQ4lPiDT2+z47fU6St2MeOHsg54NAnofbir1BZ22kQSuDfl6yyLeCUs+FDMWh9oARb34ys+2voZB6Z7ErwufzOXzcvmWKtoICO1HPejXsN9J3ureUZWkld3RGThSEk2SK8J0EdjUqBFpAYCjxKboSNM\/zhrzN0Q5QqwOJlJN5uDFCgWHO4a1IQqcIDMWh9gYhz\/OdaBYaqiJw9shpUCU+8NRP4F1dXf3zn\/\/cVZMqNcqyvLu747ojaLLTJyasZNeRvaHuZ0URvn7aRVoJge0IDCI+4WYa3n\/Bh8L8sNuOObVC+OwtPmxxjxynJGcOGAL3kKkqR0ZgEA51CCbtEr7VXBqfu+xUHwb\/dDolCbM6z9d\/e2jS3Gdn6z\/baH3aSa5nq\/Wien5VhK\/nBpJ6QqASgUHEpyp+hpvs+fn5X3\/95Y6NcR\/6tlwKtezbYa6Ww4QBCuB1LS7ZiQS87smy7OnTp9yrjmd63ApBQNHiZDL5888\/z8\/P8a1KFrMCaRurJDLDHKo6m80obdVunudACf1iMTTUXBlWnM1mqJVlGQ\/jIaruUpZlmGp1+V2c30O4+pkYhEMdAl09zZrNZhzbfD\/rNv+x9ZAkWa90V22tq6ursixrNNlJDUpmomaKrqHkKuXZxHATInzDtZ00HzsCg4hPVYQP92W76Ae0I5wSIxEhNaHhOa+AMqhrOZ\/Nz\/OcMQnMD+VXPAkTFagIzoe0ZUI2Xa\/k5q66\/o8lw+NtEPYgdrFYXFxcLJfLmh5ZBWyavUCC4cp2gWoAOnsJO2OihoiKAqQHKkAL9ioxCIc6BLEamoVh0JzwcaBCH0sQqzhiWZY8t6lGk4a0rAoHe0yUK9NQMm5NnLx0Qgb9U4Rv0OaT8qNGYBDxqZ7wIcCQWETNSdKGRPRGbCVE0zbTBqQwbpGBTadTR6FsRacqlWS+zUHrlgjyXTam5chEWd0qvDUdlW9rUSwS7hJVdfn8yYRDgPlIuA7yqmu9tz8H4VCHoFdDs3YlfI7huZ8cUVbboiju7u6QU6NJQ1pmJdt0zU7b5pJ3fSlsFehzWoSvz9aRbkKgDoFBxCdEffIb9gchwU6nuVkoVxJXw60elmrU8DNH7MhF5vM5GB6aYzEWqKI4VI8V+Y44zKFYV4v7jjmzwuZwa67pEUvi8OtwyZTtgmvXXcLPPM+j+faobtuoTUc76KQ5HXr4cxAOdQhuNTTrQMLnBoD7CZ255MA+GIRev5WW4RkSQtyNBasjqiDaKpkVRfgIhRJCQAj0AoGhxKetu3TrmYGbLbB3bcYVKyGadkJYxuUXRXF2drZY\/7U4w+da4ehZLpfPnz\/HSkFwvuY9cnzL0U17lc0xwe4jhz+ZqMm3ZZiOdpBX2W7PE0NxqL1hbJHwcaBCGbdyrr6hjtbwrYhgdPqfcNlbh52SdANVr3SJmBJCQAj0BYGhxCe7XA\/Y2SV0pCZVN2tHJnB3xjtQLsuzt+xoGjpwMo8ynW58so8KoarhtARmHWw+mmCnLOtdCb+9vQXbw05GNIfVhHY5XcMZPkzPsHeQD5lUwI5aXKK2DISuCosx4RCw+WEHnTSrQD\/TQ3GovdGjoaMSZrMZxh6d4v7+\/ubmxlqZFcNMbm4NL7EWEzWMcCc1KHA+n9vZvuvr63CPcEPJTfRnu8NK6JXusOwlbYXABwSGFZ+4Q9adtAFiFH1BAx7DF0B878l3Ot9+++0XX3yBzRxPnz7NsuzFixf44P5kMnn16hX2\/64k5HmOMAZp2KWLzbw2375ihlgKcQLJlqJK2rdXUNu2Mp1O379\/z\/2\/mPCAYg17FCrj5L99+5Z9t4EQowchzQHrDGHLPHnyZCsaOynwYRD3KTUsh9oJOWtNPhg4CRy0k8nkzZs38KNff\/2VJ1i4h4c8z93Qoo\/TOzAq6LlssZ7wYWQ2VMO6jxvSbA6Jhh0kc3XVE\/i5wSeBntR3wR7\/UpYlA4aLPfVCDrnKoeZ8ppXvdXG64hANo3W7k8zmWmyComrW7bLdBBIJx6cErNPnLiQ8h3EI7CN3qF1vm\/bDy4fA7uruqoarXvOziWQwVBLWGmlDvDQKwmdXG8Ccdk+ce7vUhRVXTYDn8Q0UWwFHOeSUp8MlUJkwQQoVXmorp60mLA6wcsit29K5J3JGHp96YoUhqiHCF7XamB1qvyFhY2sU0l0z91OjSStNJCcfONInfO4pxC404SgJt\/7x0uGJqicG7lFvMhCjahwuISr2OJlUvsXmLJJVsLfY3MlFjTk+nRz84Spg328k\/1C0k5nG7FC4ee4xHuw5tjuhHS28txpRaTaziWR+LNNWTCmdPuHj7BqXG7tlB3ypGua3YmnLQijQTmtFC7BkVeJwCVWSj5BvlW+xOYdkwksxANqY41OLw0aihIAcSmNgDAgkTvhCBrDqcPgQg2Ln5+fv37\/HAlVs+cmyzK425Tq8KDXElBIAZQH7MM2lsmgOJafT6bt37\/ABCMhv0uJWCVXHVUW7EM3E6MclLGhYUWfqzGOa6CRh9\/mOFYc78ZgEp\/yLFy\/4RjsUwsKhRag2EUNhLr9wP6lqMgkRvmRMqY70AQE5VB+sIB26QwARPOuugdNKdisMQL9CwgeeYb+GAApiJ6I42VtFI1b8A5JRgJwvWt5mIr1ri1sloABYJjhQtAvRTFqNrdjzang8Dotho2LYfasDyliQoRWbqBfi8KFlrY2sKH7SNjS3VXvQacWnQZtPyvcNATlU3ywifdpFIHHCZz+uyM25IQMA4cNEkSUNTCOxAWv9H\/kc7EH+gZ\/28\/0UYi1nM6PprS1Ga\/G1tfs4PvNdF5q3Al7les0eVXXfKslX5043lmkihIXZtM2xaRTgZ9VYPqWE4lNK1lRfTo6AHOrkJpACnSIwLsIHQhCyFstmLGlg2nGR0CSW4YHZ8KQjCrG1bGY0vbXFaC0SO0eqnErUZKdW+Ho6ZMxV3bdKUjd3ghPLNBHCwpSGERxOFqKPIny0tRJCQAjUIyDCV4+Prg4dgcQJX0hotu7SDSkFjqoMTy6ytrcvFsGucEATeQkXlqFWtBVb2Am0bTWUYJuws2tW1E6toGIUQCcnej6V7Z3VjekmQmxhUGrmWPlQFRO3IT21CAw6rfg0aPNJ+b4hIIfqm0WkT7sIJE74LBsAcHa5HnKwF4GEzFaxaUt0Vvm3t7fWEk6snVhyPAa1IDnP86Iofv\/9d3dqJzhKfYtbJbAA9YwKjGayCoSA8l5eXoJUnZ2duVNrqrrP6hDIg32om+1+EyEUyC8aMickfPYSe5RSQvEpJWuqLydHQA51chNIgU4RSJzw4dCV8B0uN5y6kzZAEdwhQiuM3LlM9rvNNA\/4CgBli3wN6hricUxff\/31559\/vl+L2KZaJeGzzz5zylgN2YVoJjpFNLIs+\/nnn3lSU3TOzMph962E6JZnHiSFPRmhEEpwZ0m9fPmSJ2i5M7VgLyzZ5AZemimlRPLxqcZ9UrJjdAKem9DpTYd3OfrwebjYZCQk71A7WQp34\/AWilHkgksombdu3NttgYYSbJVD0hr2RA9WS3aXbtWMFPuvRKcIwOc5e9ppW0447lYnadpp0t3PhOMTzMfHEh6XmapBXUzC10OLovjss8+ePHkSfgVpv0HlWtlPSMK1EnaoXa1GuhYSvuhXGpx8+zVm9w6tLMsmEpzAvX+SXKZ669gJmfQJX9V+hZ1gUuH9EDgV4QNdiM5E7teRftZKOD7ZlQYEv9MTcdjKyRNjeFY5OchRBRJ2qGh\/t2bmee4IX1EU19fXWyve3d1x2Q\/GMyeqG0rY2kTzAqcKQ801PFrJURA+fJLt4uKirQflo5ln0A3Z93FH5l78uOCgAdyqfKrxCTdoRgjigIf1MJ8F0kgoPp3Kjqk61N54hoSPKw12mjCzi9r3k7B3F8K13YeIGnrdsRC+odtJ+guBEIFU4xMeFcKHBDCh8\/PzqoNkCBHf4+AGx0NuGGxmsxmkYYER8u1khi1JsUxwEfB0On379i2Wt7oTZVg4KoqZeG2Nn\/P13+amnGWffvqpXf\/kqlA+O+sUcPnT6RSn2mA97mQy+fPPP8\/Pz7Msm81mmIaJLrRFgVBaes\/PqToUh8quiZDwQQI8tOGjV3TGukqCG2Y8nwn5WwcthzEGLbTVExTtLsJHKJQQAgNDINX4VEX4cDcHQ8JN3B4kQ+PZ+zt3c2ObFKYlWAAJkCpEFBTgDDFLUjiWiDx\/\/ny5XHI1EopZWsnVh1FRnPBAxaovU9rWwypWpSoFbL79NqfNZygt1n\/8\/lSoua210wSPVbXP6VQdam\/MqwjfTtNm9pQmqwmGUziQ7DALz2eCl9UMWjwoQggoaVVDVpmRpEX4RmJodTNBBFKNT\/WED\/NwNTdx+\/VNplGek2eYALBCmI6WtKMHwcZOb7AuipE7RkVRJSvTSmCaiWiVqup2zy8loDB\/MuEiN\/ORqIHLtp5MOlWH2ttANYSv6gsYYVuXl5dcz+eu5nlu\/QhXOQjx03mTe2azA9i5Cb\/k7wQ6HUb1U4RvVOZWZ5NCINX4hBt0GAks06q5iWMiEA\/6PFzRBQOMAyuE6WhJN25ASVd3TzudwLkKiMJXNnniDiVQJebYoGXTVClapao6JbiJQ+bbqT6badNREKiPbTqldKoOtbeN6gnffD6Hk9Id7ItUNJrnOf0iVIMS7CU3zOhNNj+aJsODNI5hW9g2NMK0CN8Ija4uJ4JAwvFp6y7d+ps4qCEXANlJL2t7K4RpzijYktE0lWRdFOPPqCjGISuTVSzrYma0SlX1qASnGCXbwjYd1dzWsq0nk07YofazUT3hqydz+BJquBLXahKV4IYZfzJhB6pNu0FbdeCTVWBsaRG+sVlc\/U0HgYTjk12uB4O5r3nh7h8NJ4vF4tmzZ+GWAvIzBInb29uqEBKWtIOmKAp36oyVg1VH3P8RinJdWywW+CwZj9th3KLYaBWrEksiMzzSBvksxoSNly4dao5aUcytMsNNJ+xQ+xmFAymsXrUyjyXd7N319bV7sVslwQ5O6002P5p2buKWvdZMNFLn5BMifMmbWB1MFoHk4xM3w7qDakCJcPMK3\/zivr+5ta3\/wxYKmz+dTt+9e8fDWl69emUPknElHXe0rdtXumyRbA8fhMJOWNsFJ5+aZFn2ww8\/UM4nn3yC9Hw+d1WcSoh\/rAgFrJ52V3KWZU+ePHGn+7iTbMIWuRk5fG2XjIMl71DNLWVHFHcj2RFlB3ko1nouhmU4Jqsk2Ka5bZyZdqDaNLZeWTfBnYEVV2ok\/KwSmiCas7lFRK8pUwgIgT4joPgUtc5yuby6usIuWjdrFS1\/eCaCygmnEE6uwOEY9kGCHKoPVtBg7s4KInzdYSvJQqBbBBSfovjO53M7eWC\/zBItf3jmyUPUyRU4HMM+SJBD9cEKGszdWUGErztsJVkIdIuA4lMVvvxGMV8JVZU8PN9uUTzJO6OTK3A4hj2RIIc6uSE0mDs1gQhfp\/BKuBDoEAHFpw7BlejxISCHGp\/Nx9XjDeHTf0JACAgBISAEhIAQEAKJIzAulqveCoEkENCERBJmVCf6goAcqi+WkB7dILAhst0Il1QhIAQ6REDxqUNwJXp8CMihxmfzcfVYhG9c9lZvU0JA8Skla6ovJ0dADnVyE0iBThEQ4esUXgkXAh0ioPjUIbgSPT4E5FDjs\/m4eizCNy57q7cpIaD4lJI11ZeTIyCHOrkJpECnCIjwdQqvhAuBDhFQfOoQXIkeHwJyqPHZfFw9FuEbl73V25QQUHxKyZrqy8kRkEOd3ARSoFMERPg6hVfChUCHCCg+dQiuRI8PATnU+Gw+rh6L8I3L3uptSggoPqVkTfXl5AjIoU5uAinQKQIifHvCaw\/rBIiTyaQoirIsl8vl+fk5MmezGRuwpwTO53PmH5gINUHTLTZhNSyKYjKZzOdz2y7Oqg9zbEWmLQ7T6XSxWPCSEjshoPi0E1wqLATqERinQ9n7NgNWnucbcpBl3Z0QjViJ8EHTLBaL6XS6NYohEoVxlnKqEgxhVQUSzt\/YNOEedtc1O24wcMn5LO2zxGu5XF5cXLROcawm6O98\/dd63+lj6BSom+0gClTdIICSJXm4rVgJreucsMBxxqeEDaqunRaB0TrUfD63wQtWWCwWz549az1a0cQkdpbw2RBZHx2urq720M2FMCozkoQI3\/6GdjQL7IdcZ7lcXl1dfffdd9aRkLlcLvdvNVbTaRIr0lqebcum0UCYYxuezWYWDVzK8zzMtLWUrkJgtPGpChDlC4FDEBitQxVFMZ1O8YaKAO5K+IqiuLu7Y\/WGiTzPLeG7u7ujGpgg4KSjFVgUxfX1tc1pngbRHOcsgwhf83HiSzpyEyV89\/f30+mUA\/o4hO\/m5qZ1TsnOW29xCJRlGea4iqH3okqYz4pKVCEw2vhUBYjyhcAhCIzWoQ4nfDV3\/nqLOMLnCs\/Wfy6zLEu+ht6Dt9kQFkpOO0eEb3\/72iGOMWRfVpLbgQiC0DATrXLUkhGWZcnFE1YaS0aJkdPk4uJiuVwiM8uy169fn5+fU1pUVDTTQkOtnj59ijV8UXpnNbHVy7J0hJhXAZ1FgJeUqEdgtPGpHhZdFQL7ITBah6ohfG\/fvp1MJowjWZaFL2RwDweZQKDB5BxyojGLBqohfBBSQ+lsbKVAJmazGcPKfD5nBBThy4iREs0RIKOK+oDldlyLYDPpY5YkFUXx\/PlzzM9xjcJsNsOgrxqpVpMsyzjKUR7+CQlRUdFMi0Oe5\/QW9oWEb\/PQYP6LumgV4QsX9tmmla5BYLTxqQYTXRICeyMwWodiMLLQ8ZWujSOYXWM4YHkXm1YzCFjdhPwazldD+Gw0ZEMu4dq1V0X4LBpIb6J0eEE5WxEgUUOCq\/dQ0XI7cpr7+\/urqyv3vtUOWYiy7oGrhk1l9iraoiZlWS4WC8zwIT2dTkm\/oqKimbbvVj0n07YbamKF1MzwARySVFdLP2sQGG18qsFEl4TA3giM1qGaED7GkfC2H8YFuyIwuiOENqohfJeXl1zPx\/JhIs\/zMCaCmDKsaIYPuInwheOnaY4d9+HOA0v46A9fffXV5eUlCZ8lW3QnzIStDAMGGXVFp6LVpCxLruFzXC0qKppp5bsCVqZrl3N+7IuVg4qhZ0JImG\/rKh1FYLTxKYpGDzPpy1kWeU7rocIjV2m0DhXeyRGzsEvX3vMZy9wEhy3jGJ6LIG6MVRG+PM+jccRVx1RCNHxohi\/ESoQvxKRpjnMSO7zwWRY3mYe7P5856AbWVdg2N7S6VljAJqrKOMnRYtHMGuFWZljX5qAkmSseucL1HyFXtq0rXYPAaONTDSa6JAT2RmC0DoV7teNw8\/kc0cre80n4HBuzZWwUwCzA2dlZ1SdUooRvFSudMjU2raKGNiJrhg8AivDVDKQtl9ywxojn4gb7PSEKskOQC+Ocq1xeXsKpzs7OMKFN8of829tbCkTCacKrkGw9JyoqmkkheOVKombbAoW1no+raPH29nax\/mNH+Gqbzm9XBLJFJRoiMNr41BAfFRMCOyEwZodyD972Pm8jlHtVSngZa4qi+Pe\/\/+22CUZn4FDXxkTk4Kv+lHx9fV3zYrdmnd9sNkM4Zty5v7+\/ublx3WFDY0iI8O1pZW5rtZsk+Abnyy+\/3CCbZZYP2fe8GHbY8PH06VO89IGboS6JGsYrMkkoqXdUE75dRS36W1RUNJPy7UeksyzDLl30mn3Eg2CVJm4pBjf8ZlkWdse2q3Q9AmOOT\/XI6KoQ2AOBkTuUvTPzCZ9Teu5uH8KL+z9igY0pjD6uCiOg3fVodUCLViDSNkoix0nGT8ajyWTy5s0bnN7x66+\/8hgPRtho9SQzN0ZMsm\/qVB8QWCwWV1dXfdAkPR1GHp\/SM6h6dFoE5FBR\/Ac6JeYmGqJdG1umCN\/YLH7s\/uLF7rFbHUd7ik\/jsLN6eSQE5FBRoIdI+IaocxT8djNF+NrFU9I+QmA1P5\/n+cr3wnWHH5XTj70QUHzaCzZVEgJxBORQIS5cp7TiCgN6BwrCNyCFQ+S7yBHh6wJVyXxAgEsoso8XMgqdthBQfGoLSckRAqtTjuRQGgZpIyDCl7Z91buUEVB8Stm66tvREZBDHR1yNXhUBET4jgq3GhMCLSKg+NQimBIlBORQGgNpIyDCl7Z91buUEVB8Stm66tvREZBDHR1yNXhUBDaET\/8JASEgBISAEBACQkAIJIzA\/wH6aFvIId5GYwAAAABJRU5ErkJggg==\"\u003e\u003cbr\u003e2. Optional. If the RNA template is GC-rich or contains secondary structures, mix gently, centrifuge briefly and incubate at 65 °C for 5 min. Chill on ice, spin down and place the vial back on ice.\u003cbr\u003e3. Add the following components in the indicated order:\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 637.604px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 383.863px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 253.009px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eReaction Buffer (5X\u003cfont\u003e)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e4 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNase Inhibitor (40 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e0.\u003cfont\u003e5\u003c\/font\u003e \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003edNTP Mix (10mM each)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\n\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eM-MLV Reverse Transcriptase \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e(200 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eTotal Volume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e20 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e4. Mix gently (gently with a pipette or gently with a vortex mixer at the lowest speed), then centrifuge the precipitated liquid.\u003cbr\u003e5. If Oligo(dT)18 or gene-specific primers are used, incubate at 42℃ for 10-60min. If random hexamer(random hexamer) is used as primer, incubate at 25℃ for 10min, and then incubate at 42℃ for 10-60min. Reverse transcription of cDNA below 3kb can be done for 10min, reverse transcription of 3-6kb cDNA can be done for 30min, and reverse transcription of cDNA above 6kb is recommended for 60min. When random hexamer(random hexamer) is used as primer and subsequently used for qPCR, 10min is sufficient for subsequent detection of genes of any length.\u003cbr\u003e6. Terminate the reaction by heating at 70°C for 5min. The reverse transcription reaction product can be directly used in PCR applications or stored at -20°C for less than one week. For longer storage, -70°C is recommended.\u003cbr\u003eIII. PCR Amplification of First Strand cDNA\u003cbr\u003eThe product of the first strand cDNA synthesis can be used directly in PCR or qPCR. The volume of first strand cDNA synthesis reaction mixture should not comprise more than 1\/10 of the total PCR reaction volume. Normally, 2 μL of the first strand cDNA synthesis reaction mixture is used as template for subsequent PCR in 50 μL total volume.\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e1. Use DEPC to dispose of all equipment used in the study, or purchase equipment that has been proved to be nucleic acid-free. Wear gloves during the study and change them frequently to avoid RNA enzyme contamination.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e2. Ensure that there is no RNA enzyme contamination in the reagents used.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e3. Keep the kit tightly sealed. During the reverse transcription process, all tubes must be securely fastened.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e4. Purified RNA must be free of salt, metal ions, ethanol and phenol, which will interfere with the first strand cDNA synthesis reaction. Trace contaminants can be removed by precipitation of RNA with ethanol.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e5. In order to ensure the effective retrotranscriptional reaction, high-quality RNA templates need to be used. \u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e6. When the template content is low or the subsequent PCR amplification fragment is too long, the reverse transcription time can be appropriately extended.\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50Rxns","offer_id":41519868837963,"sku":"UA070077-50Rxns","price":85.0,"currency_code":"USD","in_stock":true},{"title":"50Rxns*5","offer_id":41519868870731,"sku":"UA070077-50Rxns*5","price":335.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/d3d6a297-3e26-415a-9023-06fc3b4983f8.png?v=1787958082"},{"product_id":"startscript-ii-1st-strand-cdna-synthesis-kit","title":"StartScript® II 1st Strand cDNA Synthesis Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePhysical Appearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eLiquid\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e[1] Roth, M J ,  N. Tanese , and  S. P. Goff . \"Purification and characterization of murine retroviral reverse transcriptase expressed in Escherichia coli. \" Journal of Biological Chemistry 260.16(1985):9326.\u003cbr\u003e[2] Kotewicz, Michael L , et al. \"Isolation of cloned Moloney murine leukemia virus reverse transcriptase lacking ribonuclease H activity.\" Nucleic Acids Research 16.1(1988):265-277.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eThe StartScript II cDNA First Strand Synthesis Kit contains two optimized mixes: StartScript II Enzyme Mix and StartScrip\u003cfont\u003et\u003c\/font\u003e II Reaction Mix. The enzyme mixture contains StartScrip\u003cfont\u003et\u003c\/font\u003e II reverse transcriptase and mouse RNase inhibitor; the reaction mixture contains dNTP and optimized buffer. StartScript II reverse transcriptase is a recombinant M-MLV reverse transcriptase that reduces RNase H activity and improves thermal stability. StartScript II reverse transcriptase synthesizes first-strand cDNA at higher temperatures compared to wild-type M-MLV reverse transcriptase. The enzyme has an activity temperature of up to 48°C and has the characteristics of high specificity and high cDNA yield.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eThe kit also offers two optimized reverse transcription primers with nuclease-free water contaminated water. The Oligo-dT primer [d(T) 23 VN] can anneale with the start of the polyA tail. The optimized random primer mix can be randomly and continuously paired with the entire RNA template, including mRNA and polyA-free RNA. The synthesized first-strand cDNA product can be up to 10 kb in length.\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 709.34px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 300.6px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eComponents\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 216.009px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070076-50 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 192.002px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070076-250 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eStartScript II Reaction Mix (2X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e600 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 x 600 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eStartScript II Enzyme Mix (10X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 x 100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003eRandom Primer Mix (60 μM)\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e110 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 x 110 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOligo(dT)23 VN* (50 μM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e110 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 x 110 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase-Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 x 1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e* V = A, G, or C; N = A, G, C, or T\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eIf denaturation of template RNA is desired, use the following protocol.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1. Mix RNA sample and primer in a sterile RNase-free microfuge tube.\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 680.34px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 243.6px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eNumber\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 270px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 166px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1. Template RNA\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e(One of the three is optional)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eTotal RNA\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOr poly(A) RNA\/mRNA\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOr specific RNA\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u0026lt; 5 μg\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u0026lt; 1 μg\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u0026lt; 0.5 μg\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2. Primers\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e(One of the three is optional)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOligo(dT) 23 VN (50 μM)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOr Random Primer Mix (60 μM)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOr Specific Primer (1 μM)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 µl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e3. H2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUp to 8 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2. Denature sample RNA\/primer for 5 minutes at 65°C. Spin briefly and put promptly on ice.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e3. Add the following components\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 676.34px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 367.6px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eStartScript II Reaction Mix (2X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 308.007px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.4px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eStartScript II Enzyme Mix (10X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.4px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e4. Incubate the 20 μl cDNA synthesis reaction at 42°C for one hour. If Random Primer Mix is used, an incubation step at 25°C for 5 minutes is recommended before the 42°C incubation.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5. Inactivate the enzyme at 80°C for 5 minutes. The cDNA product should be stored at –20°C. In general, the volume of cDNA product should not exceed 1\/10 of the PCR reaction volume.\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1. Use DEPC to dispose of all equipment used in the study, or purchase equipment that has been proved to be nucleic acid-free. Wear gloves during the study and change them frequently to avoid RNA enzyme contamination.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e. Ensure that there is no RNA enzyme contamination in the reagents used.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e3\u003c\/font\u003e. In order to ensure the effective retrotranscriptional reaction, it is necessary to use high-quality RNA templates.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e4\u003c\/font\u003e. Poly (A)+ mRNA does not need to be isolated from total RNA, but using Poly (A)+ mRNA as a template can improve the yield and purity of the final product. The amount of RNA used, the total RNA is less than 5 μg; Poly (A)+ RNA is less than 1 μg; Specific RNA is less than 0.5 μg.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5. The cDNA products should be stored at -20°C.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e6. Enzymes should be placed on ice during experimental operation, and stored at -20°C after the experiment.\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50Rxns","offer_id":41519868903499,"sku":"UA070076-50Rxns","price":85.0,"currency_code":"USD","in_stock":true},{"title":"50Rxns*5","offer_id":41519868936267,"sku":"UA070076-50Rxns*5","price":335.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/03e55ad1-d932-4fb0-beb2-86bd4f109ebb.png?v=1787958084"},{"product_id":"phi29-polymerase-rca-kit","title":"Phi29 Polymerase RCA Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eThe Phi29 polymerase RCA (Rolling Ring Amplification) kit contains all the reagents needed to achieve sensitive and efficient rolling ring amplification (RCA) , an isothermal amplification method that continuously amplifies circular DNA by producing long repeat copies of circular sequences. The kit contains phi29 DNA polymerase, dNTPs and random primers for exonuclease resistance (containing phosphorothioate bonds), ensuring the versatility of amplified circular DNA sequences.  The starting material can be purified circular DNA (single or double stranded) or use liquid medium culture, agar plate colonies and glycerol directly without additional DNA extraction.  RCA products can be used directly in downstream applications such as DNA sequencing, cell free DNA enrichment, cell-free protein expression, and DNA biosensors.\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 752.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eContain\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 179.053px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070120-50 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 175.002px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070120-250 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003ePhi29 DNA Polymerase (10 X) \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 X RCA Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e500 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 500 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eExonuclease-Resistant Random Primers (500 µM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eDeoxynucleotide (dNTP) Solution Mix (10 mM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e1. Prepare the reaction without adding enzymes, as shown in the table below. Mix thoroughly by pipetting or vortexing, but it should be gentle. Centrifuge briefly to collect the solution to the bottom of the tube.\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 656.41px; height: 311.088px;\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 33.0347px;\"\u003e\n\u003ctd style=\"width: 353.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 148.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 154.002px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eFinal Concentration\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 51.7245px;\"\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 X RCA Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e4 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 X\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 51.7245px;\"\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eExonuclease-Resistant Random Primers (500 µM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e50 µM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 51.7245px;\"\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eDeoxynucleotide (dNTP) Solution Mix (10 mM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 mM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 51.7245px;\"\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eCircular DNA Template\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eX µl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e0.01-50 ng\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 51.7245px;\"\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUp to 18 \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 X\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e2. Incubate in a thermal circulator with the lid set at 100°C and incubate at 95°C for 3 minutes. Then allow the sample to cool to room temperature.\u003cbr\u003e3. Place samples on ice and add 2 µl of phi29 DNA Polymerase to each sample. Mix thoroughly, and gently, by pipetting or vortexing. Centrifuge briefly to collect solutions to the bottom of tubes.\u003cbr\u003e4. Incubate in a thermocycler with the lid set at ≥ 75°C, incubate at 30°C for 2 hours, followed by 10 minutes at 65°C to inactivate the DNA polymerase. The RCA products can be kept overnight at 4°C or for long term storage at -20°C\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e1. RCA amplification is the amplification of circular templates using the characteristics of Phi29 DNA polymerase chain displacement activity and continuous synthesis ability. Therefore, its optimal substrate is circular ssDNA or dsDNA templates (C-DNA). In addition, the linear template (L-DNA) with larger molecular weight can also be continuously synthesized and amplified, such as genomic DNA, in which the amplification ratio is weaker than that of CDNA.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e2. The RCA products may be viscous due to the high yield of high molecular weight DNA. Double dilution with nuclease-free water is recommended before using any downstream applications..\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50Rxns","offer_id":41519879913547,"sku":"UA070120-50Rxns","price":275.0,"currency_code":"USD","in_stock":true},{"title":"50Rxns*5","offer_id":41519879946315,"sku":"UA070120-50Rxns*5","price":1165.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/5ab892c8-2ab8-4f00-b0e4-66b2a22c4cea.png?v=1788012127"},{"product_id":"alexa-fluor-488-antibody-labeling-kit","title":"iFluor® 488 Antibody Labeling Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e12 months from date of receipt \/ reconstitution, -20 °C as supplied\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eiFluor® 488 antibody labeling kit is used for iFluor® 488 labeling and conjugation of antibodies.\u003c\/p\u003e\u003c\/div\u003e","brand":"Starter","offers":[{"title":"500μg","offer_id":41677885210699,"sku":"S0E1002-500μg","price":350.0,"currency_code":"USD","in_stock":true},{"title":"1mg","offer_id":41677885243467,"sku":"S0E1002-1mg","price":650.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/0e2d6a32-54ae-4fc0-a7b6-e102e98161ae_b4403a7c-d374-4da6-b2bc-dbf52a59feeb.jpg?v=1790157610"},{"product_id":"startscript-ii-1st-strand-cdna-synthesis-kit-with-dnase-i","title":"StartScript® II 1st Strand cDNA Synthesis Kit, with DNase I","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePhysical Appearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eLiquid\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e[1] Roth, M J ,  N. Tanese , and  S. P. Goff . \"Purification and characterization of murine retroviral reverse transcriptase expressed in Escherichia coli. \" Journal of Biological Chemistry 260.16(1985):9326.\u003cbr\u003e[2] Kotewicz, Michael L , et al. \"Isolation of cloned Moloney murine leukemia virus reverse transcriptase lacking ribonuclease H activity.\" Nucleic Acids Research 16.1(1988):265-277.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 789.604px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 204.85px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070078-10 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 187px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070078-50 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eDNase I, RNase-free (2 U\/μL)  \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10X Reaction Buffer with MgCl2 for DNase I   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e50 mM EDTA   \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eM-MLV\u003cfont\u003e(\u003c\/font\u003eH-) Reverse Transcriptase (200 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e5\u003c\/font\u003e0 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5× Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e40 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase Inhibitor (40 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e25 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003edNTP Mixture (10 mM each)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e5\u003c\/font\u003e0 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOligo(dT)18 Primer (50 μM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e5\u003c\/font\u003e0 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRandom Hexamer Primer\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e(50 μM)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e20 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.014px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.025px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003eI. Removal of genomic DNA from RNA preparations\u003cbr\u003e1. The reaction system is formulated as follows:\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 729.604px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 365.863px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 363.007px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNA\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10X Reaction Buffer with MgCl2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eDNase I, RNase-free (2 U\/μL)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNase Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eTo 10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e2. Incubate at 37 °C for 10 min.\u003cbr\u003e3. Add 1 μL 50 mM EDTA and incubate at 65 °C for 10 min. RNA hydrolyzes during heating with divalent cations in the absence of a chelating agent. \u003cbr\u003e4. Use the prepared RNA as a template for reverse transcriptase.\u003cbr\u003eII. First Strand cDNA Synthesis\u003cbr\u003e1. Set up the reverse transcription reaction according to the following table\u003c\/p\u003e\n\u003cp\u003e\u003cimg class=\"img-fluid\" src=\"data:image\/png;base64,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\"\u003e\u003cbr\u003e2. Optional. If the RNA template is GC-rich or contains secondary structures, mix gently, centrifuge briefly and incubate at 65 °C for 5 min. Chill on ice, spin down and place the vial back on ice.\u003cbr\u003e3. Add the following components in the indicated order:\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 635.604px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 396.863px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 238px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eReaction Buffer (5X\u003cfont\u003e)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e4 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNase Inhibitor (40 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e0.\u003cfont\u003e5\u003c\/font\u003e \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003edNTP Mix (10mM each)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\n\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eM-MLV(H-)Reverse Transcriptase \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e(200 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eTotal Volume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 595.532px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e20 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e4. Mix gently (gently with a pipette or gently with a vortex mixer at the lowest speed), then centrifuge the precipitated liquid.\u003cbr\u003e5. If Oligo(dT)18 or gene-specific primers are used, incubate at 42℃ for 10-60 min. If random hexamer(random hexamer) is used as primer, incubate at 25℃ for 10min, and then incubate at 42℃ for 10-60 min. Reverse transcription of cDNA below 3 kb can be done for 10min, reverse transcription of 3-6 kb cDNA can be done for 30 min, and reverse transcription of cDNA above 6 kb is recommended for 60 min. When random hexamer(random hexamer) is used as primer and subsequently used for qPCR, 10min is sufficient for subsequent detection of genes of any length.\u003cbr\u003e6. Terminate the reaction by heating at 70 °C for 5 min. The reverse transcription reaction product can be directly used in PCR applications or stored at -20 °C for less than one week. For longer storage, -70 °C is recommended.\u003cbr\u003eIII. PCR Amplification of First Strand cDNA\u003cbr\u003eThe product of the first strand cDNA synthesis can be used directly in PCR or qPCR. The volume of first strand cDNA synthesis reaction mixture should not comprise more than 1\/10 of the total PCR reaction volume. Normally, 2 μL of the first strand cDNA synthesis reaction mixture is used as template for subsequent PCR in 50 μL total volume.\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e1. Use DEPC to dispose of all equipment used in the study, or purchase equipment that has been proved to be nucleic acid-free. Wear gloves during the study and change them frequently to avoid RNA enzyme contamination.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e2. Ensure that there is no RNA enzyme contamination in the reagents used.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e3. Keep the kit tightly sealed. During the reverse transcription process, all tubes must be securely fastened.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e4. Purified RNA must be free of salt, metal ions, ethanol and phenol, which will interfere with the first strand cDNA synthesis reaction. Trace contaminants can be removed by precipitation of RNA with ethanol.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e5. In order to ensure the effective retrotranscriptional reaction, high-quality RNA templates need to be used. \u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e6. When the template content is low or the subsequent PCR amplification fragment is too long, the reverse transcription time can be appropriately extended.\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50Rxns","offer_id":41678281867339,"sku":"UA070078-50Rxns","price":100.0,"currency_code":"USD","in_stock":true},{"title":"50Rxns*5","offer_id":41678281900107,"sku":"UA070078-50Rxns*5","price":420.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/90045ea3056d469f8fcf26201a8d7736.png?v=1787958079"},{"product_id":"t7-high-yield-rna-synthesis-kit","title":"T7 High Yield RNA Synthesis Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e[1] Athanasios D , Kanchana R ,Hobert Elissa M.Moore Melissa J.Rabideau Amy E.An engineered T7 RNA polymerase that produces mRNA free of immunostimulatory byproducts[J].Nature biotechnology, 2023, 41(4):560-568.\u003cbr\u003e[2]Skinner, and  M. G. . \"Promoter Binding, Initiation, and Elongation By Bacteriophage T7 RNA Polymerase A SINGLE-MOLECULE VIEW OF THE TRANSCRIPTION CYCLE.\" Journal of Biological Chemistry 279.5(2004):3239-44.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 715.34px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 295.6px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 209.009px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eUA070079-50 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 210.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eUA070079-100 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eT7 RNA Polymerase (50 U\/μL) \u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eRNase Inhibitor (40 U\/μL) \u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e25 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eDNase I, RNase-free (2 U\/μL) \u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e10×Transcription Buffer\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eATP (25 mM)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eUTP (25 mM)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eGTP (25 mM)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eCTP (25 mM)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eControl Template (0.1 μg\/μl)\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e20 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eRNase Free dH2O\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 ml\u003cfont\u003e×\u003c\/font\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e1. DNA template preparation\u003cbr\u003eLinearized plasmids with double-stranded T7 promoters or PCR amplification products can be used as T7 High Yield RNA Synthesis Kit in vitro transcription templates, which can be dissolved in TE buffer or RNase free H2O.\u003cbr\u003eT7 promoter sequence: TAATACGACTCACTATAG*GG (Note: G* is the first base of RNA transcription).\u003cbr\u003eA. Plasmid template\u003cbr\u003eInsert the target DNA to the plasmid vector containing the T7 promoter, and then treated with restriction enzymes, purified after completely linearized.\u003cbr\u003eNote: \u003cbr\u003e1. The circular plasmids have no effective termination; RNA products of different lengths will be transcribed. In order to obtain a specific length RNA, the plasmid must be completely linearized.\u003cbr\u003e2. The restriction enzyme selected for plasmid linearization needs to be on the right side of the promoter region, downstream of the inserted DNA fragment, and has no recognition site in the inserted DNA fragment. The restriction enzyme should be capable of forming 5' sticky ends or smooth ends.\u003cbr\u003e3. In order to avoid the influence of protein and salt ions on the system, the plasmid is recommended to be purified when used as a template for in vitro transcription after linearization.\u003cbr\u003eB. PCR product template\u003cbr\u003eThe PCR product with T7 promoter can be used as an in vitro transcription template. First, add the T7 promoter sequence (TAATACGA CTCACTATAGGG) to the 5' end of the upstream primer sense strand; next, the T7 promoter DNA template is amplified under the action of high-fidelity enzyme; then transcription is performed. PCR products can be used directly as templates without purification, but higher RNA output will be obtained after purification.\u003cbr\u003eNote: \u003cbr\u003e1. The specificity and concentration of the PCR product must be confirmed by electrophoresis when used as a template. Add 2-5 μL of PCR product into the 20 μL reaction system.\u003cbr\u003e2. In order to obtain more high-quality RNA, the PCR product should be recovered by gel and used as a template for vitro transcription.\u003cbr\u003e\u003cbr\u003e2. In vitro RNA transcription   \u003cbr\u003eA. Thawing reagents\u003cbr\u003eThe T7 RNA polymerase and RNase Inhibitor were centrifuged briefly and placed on ice. Thaw 10× Transcription Buffer and ribonucleotides (ATP, CTP, GTP, UTP), mix and centrifuge to the bottom of the tube, place 10× Transcription Buffer at room temperature, and place 4 types of ribonucleotides on ice.\u003cbr\u003eB. Assembly transcription reaction at room temperature\u003cbr\u003ePrepare the reaction system according to the following system:\u003c\/p\u003e\n\u003ctable style=\"width: 769.34px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 291.6px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 222.002px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 255px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eFinal concentration\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e10 × Transcription Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 ×\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eATP (25 mM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e2.5 mM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eUTP (25 mM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e2.5 mM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eCTP (25 mM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e2.5 mM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eGTP (25 mM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e2.5 mM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eTemplate DNA\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003ex\u003c\/font\u003e \u003cfont\u003eμl \u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e5-25 \u003cfont\u003eng\u003c\/font\u003e\/μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eRNase Inhibitor\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e0\u003c\/font\u003e.5 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e-\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eT7 RNA Polymerase\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e1 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e-\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003eRNase Free dH2O\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003eUp to 20 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:10.5000pt\"\u003e\u003cfont\u003e-\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003eC. Incubate at 37°C for 2 hours\u003cbr\u003eMix the above reaction solution, briefly centrifuge to the bottom of the tube, and incubate at 37°C for 2h. If the transcript length is less than 100 nt, increase the reaction time to 4-8h.\u003cbr\u003eD. DNase I treatment (optional)\u003cbr\u003eAfter the reaction is complete, add 2 μl of DNase I (RNase free) to each tube and incubate at 37°C for 30 mins to remove the template DNA.\u003cbr\u003e\u003cbr\u003e3. Product purification \u003cbr\u003eAfter electrophoretic analysis and purification, the synthesized RNA can be used for downstream experiments.\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e1. As spermine components are contained in 10×Transcription Buffer, it will precipitate with template DNA at low temperature, so preparation of reaction solution should be carried out at room temperature, the order of component loading should be adjusted, the system should be calculated, and water, buffer and NTP should be added first, and template and enzyme should be added finally.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e2. Perform the reaction in a PCR machine with the hot lid open to prevent the reaction solution from evaporating for a long time.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e3. The reaction product may have a white precipitate. This is free pyrophosphate and magnesium ions produce the magnesium pyrophosphate in the reaction, won’t affect the subsequent experiments. You can add some EDTA to clear it. If the addition of EDTA affects subsequent experiments, the supernatant can also be recovered by centrifugation.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e4. The reagents and containers should be without RNase contamination.\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50Rxns","offer_id":41678289272907,"sku":"UA070079-50Rxns","price":225.0,"currency_code":"USD","in_stock":true},{"title":"100Rxns","offer_id":41678289305675,"sku":"UA070079-100Rxns","price":400.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/1fcf615a95a8441b84379e87af87c436.png?v=1787968977"},{"product_id":"fast-pngase-f-kit","title":"Fast PNGase F Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003ePNGase F (N-Glycosidase F) is an asparagine amidase produced by Elizabethkingia miricola. PNGase F releases intact N-glycans by hydrolyzing the amide bonds of asparagine side chains (Asn-X-Ser\/Thr, where X ≠ Pro), while converting Asn to aspartic acid (Asp). Unlike other glycosidases (such as Endo H), PNGase F can cleave all types of N-glycans (high-mannose, hybrid, and complex oligosaccharides), but it does not act on O-linked glycans or plant\/insect N-glycans modified by core α-1,3-fucosylation. This enzyme is of significant importance in glycoprotein research, quality control in biopharmaceuticals, and proteomics. To enhance the convenience of experiments, we have developed the Fast PNGase F Kit, which simplifies the experimental procedures and reduces the reaction time to less than 20 minutes, thereby significantly improving experimental efficiency.\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 576.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 169.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eConstituent\u003c\/font\u003e\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 209.005px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070119-50 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 197.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070119-100 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003ePNGase F \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e 100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5*Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e200 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e400 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1. Two \u003cfont\u003e-Step\u003c\/font\u003e deglycosylation\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e). Add the following components to one tube on ice:\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 617.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 207.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 180.007px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 229.007px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eFinal Concentration\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eGlycoprotein\u003c\/font\u003e\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e-\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e0 μ\u003cfont\u003eg\/μl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 ×\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003ed\u003cfont\u003ed\u003c\/font\u003eH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUp to 10 \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e-\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e). Denaturation: heating to 80℃ for 2 min \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e3\u003c\/font\u003e). Deglycosylation: immediately after the reaction was completed, it was cooled on ice, and 1 μl of PNGase F was added and incubated at 50ºC for 10 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2. \u003cfont\u003eOne\u003c\/font\u003e -Step deglycosylation\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e). Add the following components to one tube on ice:\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 614.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 206.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 192.002px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 215.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eFinal Concentration\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eGlycoprotein\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e-\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e0 μ\u003cfont\u003eg\/μl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5* Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 ×\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003ePNGase F \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e1\u003c\/font\u003e \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e-\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003ed\u003cfont\u003ed\u003c\/font\u003eH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUp to 10 \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e-\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003e2\u003c\/font\u003e). Deglycosylation: 1 μl of PNGase F was added and incubated at 50ºC for 10 minutes.\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003eTwo-step enzyme digestion is recommended to ensure higher enzyme digestion efficiency. \u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003eThis product is for scientific research purposes only. \u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003eFor your safety and health, please wear a lab coat and disposable gloves.\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50Rxns","offer_id":41678326956107,"sku":"UA070119-50Rxns","price":455.0,"currency_code":"USD","in_stock":true},{"title":"100Rxns","offer_id":41678326988875,"sku":"UA070119-100Rxns","price":800.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/1430bd63fae345ad9e499fa044505ac2.png?v=1788008626"},{"product_id":"fast-ligation-kit","title":"Fast ligation Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eDNA ligases catalyze the formation of phosphodiester bonds at single-stranded DNA breaks in double-stranded DNA in vivo, and T4 DNA Ligase is a type of DNA ligase. This kit contains Fast Ligase and the optimized Fast Ligation Reaction Buffer (2X) which can be used as a simple system for rapid DNA ligation reactions. The ligation of sticky-end or flat-end DNA insert fragments to plasmid vectors can be efficiently accomplished within 5-10 min at room temperature (25℃), and the ligation efficiency of Fast Ligation is equivalent to that of conventional ligation with T4 DNA Ligase for 1 hour. The ligated product can be directly used in subsequent transformation experiments without purification.\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 580.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 258.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eContain\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 162.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070123-30 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 159.009px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070123-150 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eFast Liga\u003cfont\u003ese\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e30 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e 150 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eFast Ligation Reaction Buffer(2X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1\u003cfont\u003em\u003c\/font\u003el\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 1 \u003cfont\u003em\u003c\/font\u003el\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e1. Prepare the following mixture in a Nuclease-Free centrifuge tube:\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 409.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 278.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 130.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eFast Ligation Reaction Buffer (2X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eVector\u003c\/font\u003e DNA\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eX μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eI\u003cfont\u003ensert\u003c\/font\u003e DNA \u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eY μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eN\u003cfont\u003euclease\u003c\/font\u003e-Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUp to 20 \u003cfont\u003eμl\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eF\u003cfont\u003east\u003c\/font\u003e L\u003cfont\u003eigase\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e2. Calculation of Molarity of Ends:\u003cbr\u003eMolarity = [(µg\/µl) ÷ (base pairs x 650 daltons)] x 2 ends\u003cbr\u003e3. Blow the mixture gently to mix well and centrifuge instantaneously.\u003cbr\u003e4. Leave the mixture at room temperature (25°C) for 5-10min.\u003cbr\u003e5. Transfer 1-5μl of the reaction solution into 50μl of receptor cells for subsequent experiments, or store at -20℃.\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e1. Fast Ligase is added last.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e2. Fast Ligation Buffer should be thawed in advance at room temperature and mixed well by shaking up and down.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e3. The optimal molar ratio of vector to inserted DNA is 1:3.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e4. In the case of vector single digestion for insertion of exogenous fragments, care should be taken to dephosphorylate the vector to avoid plasmid self-association.\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e5. The enzyme product should be kept on ice during the experimental operation and stored at -20°C immediately after the experiment.\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"30Rxns","offer_id":41678328758347,"sku":"UA070123-30Rxns","price":100.0,"currency_code":"USD","in_stock":true},{"title":"150Rxns","offer_id":41678328791115,"sku":"UA070123-150Rxns","price":400.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/7789fd2286904d4e9b085619f3fc41b6.png?v=1788012135"},{"product_id":"extraction-free-one-step-rt-qpcr-kitsybr-green","title":"Extraction-free One-Step RT-qPCR Kit（SYBR Green）","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eOne-Step RT-qPCR Detection Kit (Dye-Based) without RNA Extraction\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eThe one-step RT-qPCR assay kit for direct extraction-free use (dye-based) should be stored at −20°C or lower. After first use, any remaining L reagent and S reagent can be stored at 4°C for one month, or aliquoted and stored long-term at −20°C. The cell lysis L reagent supplemented with DNase I should be freshly prepared as needed and is not recommended for storage. The RT-qPCR Mix should be aliquoted and stored to avoid repeated freeze-thaw cycles. DNase I should also be aliquoted and stored to avoid repeated freeze-thaw cycles. ROX should be aliquoted and stored to avoid repeated freeze-thaw cycles.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cspan\u003eRT-qPCR is widely used for gene expression detection and quantification. Although traditional RT-qPCR offers a one-step method, it involves RNA extraction from samples, making the process complex, subject to numerous influencing factors, and resulting in very low throughput. The EF One-Step RT-qPCR Assay Kit is an extraction-free, one-step RT-qPCR kit: after cell washing, lysis buffer is added directly to lyse the cells, and the lysate is used as the template for RT-qPCR amplification. From cell preparation to the completion of qPCR, the entire procedure can be finished in under two hours. Standardized protocols reduce both the time required and the number of variables, significantly increasing assay throughput.\u003c\/span\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan\u003eOne-step extraction-free method\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003eThe components and specifications of the test kit are shown in the following table. For each sample\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eof cell lysis volume and\u003c\/span\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL RT-qPCR\u003c\/span\u003e\u003cspan\u003ereaction system,\u003c\/span\u003e\u003cspan\u003eQ1001\u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003eQ1002\u003c\/span\u003e\u003cspan\u003ecan respectively perform\u003c\/span\u003e\u003cspan\u003e100\u003c\/span\u003e\u003cspan\u003ereactions and\u003c\/span\u003e\u003cspan\u003e1000\u003c\/span\u003e\u003cspan\u003ereactions.\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003especifications\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003ereagent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eS\u003c\/span\u003e\u003cspan\u003ereagent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eDNase I\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eRT-qPCR Mix\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eROX\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e100T\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0.5 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0.4 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1000T\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5x 10 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e500\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e4x 1ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e200\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eEF One-Step RT-qPCR\u003c\/span\u003e\u003cspan\u003eFor the dye method (\u003c\/span\u003e\u003cspan\u003eSYBR Green I\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003eto ensure detection specificity and sensitivity,\u003c\/span\u003e\u003cspan\u003eall parameters of RT-qPCR\u003c\/span\u003e\u003cspan\u003eneed to be optimized, especially primer design and cell number are crucial.\u003c\/span\u003e\u003cspan\u003eqPCR\u003c\/span\u003e\u003cspan\u003eThe best primers are located at the junction of adjacent exons; such primers can only amplify using\u003c\/span\u003e\u003cspan\u003emRNA\u003c\/span\u003e\u003cspan\u003eas the template; or the primers are located on adjacent exons, and the intron between the exons is relatively long. The amplification efficiency of primers using genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eas the template is much lower than that using\u003c\/span\u003e\u003cspan\u003emRNA\u003c\/span\u003e\u003cspan\u003eas the template, so amplification using genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eas the template can be ignored. Primer design should also aim to minimize primer dimers as much as possible. It is recommended to use professional primer design software, such as\u003c\/span\u003e\u003cspan\u003ePrimer 3, IDT PrimerQuest, Primer-BLAST\u003c\/span\u003e\u003cspan\u003efor primer design, selecting\u003c\/span\u003e\u003cspan\u003e2-3\u003c\/span\u003e\u003cspan\u003epairs of primers for testing, and choosing the optimal primer pair for experiments.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eCells need to be detected within a range of\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eto\u003c\/span\u003e\u003cspan\u003e20,000\u003c\/span\u003e\u003cspan\u003eWithin a single cell range (\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003ecell lysis\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003ereagent\u003c\/span\u003e\u003cspan\u003e, 1-400\u003c\/span\u003e\u003cspan\u003ecells\u003c\/span\u003e\u003cspan\u003e\/\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003cspan\u003e, optimize the amount of cell lysis solution added according to gene expression abundance\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003ereaction volume, recommended\u003c\/span\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003eof\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003ereaction system does not exceed\u003c\/span\u003e\u003cspan\u003e400\u003c\/span\u003e\u003cspan\u003ecells\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003ethe volume of cell lysis solution does not exceed\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003e.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eRegarding\u003c\/span\u003e\u003cspan\u003eDNase I\u003c\/span\u003e\u003cspan\u003etreatment\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003eIn many cases, such as pseudogenes, genes without introns, or unknown factors, it becomes impossible to avoid genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eamplification through primer design.\u003c\/span\u003e\u003cspan\u003eDNase I\u003c\/span\u003e\u003cspan\u003etreatment can degrade genomic\u003c\/span\u003e\u003cspan\u003eDNA,\u003c\/span\u003e\u003cspan\u003ereduce the impact of genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eamplification.\u003c\/span\u003e\u003cspan\u003eThe expression level of\u003c\/span\u003e\u003cspan\u003emRNA will also determine how much genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eamplification affects the quantification of\u003c\/span\u003e\u003cspan\u003emRNA\u003c\/span\u003e\u003cspan\u003e. For highly expressed\u003c\/span\u003e\u003cspan\u003emRNA\u003c\/span\u003e\u003cspan\u003ethe amount far exceeds the number of genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003egene copies; therefore, there is a difference between products with\u003c\/span\u003e\u003cspan\u003eRT\u003c\/span\u003e\u003cspan\u003eand without\u003c\/span\u003e\u003cspan\u003eRT\u003c\/span\u003e\u003cspan\u003ethat is\u003c\/span\u003e\u003cspan\u003e\u0026gt;10 Ct,\u003c\/span\u003e\u003cspan\u003eand due to template competition, in the\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003ereaction, genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eThe amplification probability is much lower than that of high-abundance\u003c\/span\u003e\u003cspan\u003emRNA\u003c\/span\u003e\u003cspan\u003e, so the impact of genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eamplification on gene quantification can be ignored. For genes with extremely low expression levels,\u003c\/span\u003e\u003cspan\u003emRNA\u003c\/span\u003e\u003cspan\u003e(each cell contains only a few\u003c\/span\u003e\u003cspan\u003emRNAs\u003c\/span\u003e\u003cspan\u003e) in quantity, the number of genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003egene copies is comparable; in such cases, it is necessary to rely on\u003c\/span\u003e\u003cspan\u003eDNase I\u003c\/span\u003e\u003cspan\u003eto degrade genomic\u003c\/span\u003e\u003cspan\u003eDNA,\u003c\/span\u003e\u003cspan\u003ethereby reducing the influence of genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eamplification.\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003eAt this point, the duration of cell lysis\u003c\/span\u003e\u003cspan\u003e\/DNase I\u003c\/span\u003e\u003cspan\u003etreatment can be extended to\u003c\/span\u003e\u003cspan\u003e20 minutes\u003c\/span\u003e\u003cspan\u003ein order to more effectively degrade genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003e. When measuring genes in organisms known not to contain\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eat certain stages—for example, determining the expression of influenza virus genes within cells after infection—the\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003ereagent does not need to include\u003c\/span\u003e\u003cspan\u003eDNase I\u003c\/span\u003e\u003cspan\u003e.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003eCell preparation: Cell detection should be performed within a range of\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eto\u003c\/span\u003e\u003cspan\u003e20,000\u003c\/span\u003e\u003cspan\u003ecells. For adherent cells (\u003c\/span\u003e\u003cspan\u003e96-\u003c\/span\u003e\u003cspan\u003ewell or\u003c\/span\u003e\u003cspan\u003e384-\u003c\/span\u003e\u003cspan\u003ewell), after aspirating the culture medium, wash once with pre-cooled\u003c\/span\u003e\u003cspan\u003ePBS\u003c\/span\u003e\u003cspan\u003e, and try to completely remove the\u003c\/span\u003e\u003cspan\u003ePBS\u003c\/span\u003e\u003cspan\u003ebefore proceeding according to step\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003eAdd cell lysis buffer. After centrifuging suspended cells to remove the culture medium, wash once with pre-cooled\u003c\/span\u003e\u003cspan\u003ePBS\u003c\/span\u003e\u003cspan\u003e, and try to discard as much of the\u003c\/span\u003e\u003cspan\u003ePBS\u003c\/span\u003e\u003cspan\u003eas possible, then use\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003eof pre-cooled\u003c\/span\u003e\u003cspan\u003ePBS\u003c\/span\u003e\u003cspan\u003eResuspend the cells and transfer them to a\u003c\/span\u003e\u003cspan\u003e96-\u003c\/span\u003e\u003cspan\u003ewell plate,\u003c\/span\u003e\u003cspan\u003ethen, according to step\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003eadd the cell lysis buffer\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eCell lysis buffer plus\u003c\/span\u003e\u003cspan\u003eDNase I\u003c\/span\u003e\u003cspan\u003eis prepared as follows:\u003c\/span\u003e\u003cspan\u003eDNase I\u003c\/span\u003e\u003cspan\u003eis at a concentration of\u003c\/span\u003e\u003cspan\u003e100x, 990\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003efor cell lysis\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003ereagent is added\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el DNase I\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\u003c\/span\u003e\u003cspan\u003eTo the cell culture plate prepared in step\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003e(a\u003c\/span\u003e\u003cspan\u003e96-\u003c\/span\u003e\u003cspan\u003ewell or\u003c\/span\u003e\u003cspan\u003e384-\u003c\/span\u003e\u003cspan\u003ewell) add per well\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003ecell lysis\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003ereagent\u003c\/span\u003e\u003cspan\u003e(DNAse I)\u003c\/span\u003e\u003cspan\u003e, gently pipette up and down\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003etimes, and incubate at room temperature\u003c\/span\u003e\u003cspan\u003efor 10\u003c\/span\u003e\u003cspan\u003eminutes. (If necessary, to better degrade genomic\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003e, the incubation time can be extended to\u003c\/span\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003eminutes.)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4.\u003c\/span\u003e\u003cspan\u003eAdd per well in the cell culture plate\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003ecell lysis termination\u003c\/span\u003e\u003cspan\u003eS\u003c\/span\u003e\u003cspan\u003ereagent, gently pipette up and down\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003etimes, and incubate at room temperature\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003eminutes. (This cell lysis product can be used as a template for\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003e, and it is recommended to proceed with the\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003ereaction immediately, keeping it on ice for no more than\u003c\/span\u003e\u003cspan\u003e4\u003c\/span\u003e\u003cspan\u003ehours. For short-term storage,\u003c\/span\u003e\u003cspan\u003e-20\u003c\/span\u003e\u003cspan\u003e℃ storage\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003eweek; for long-term storage, it can be kept at\u003c\/span\u003e\u003cspan\u003e-80\u003c\/span\u003e\u003cspan\u003e℃)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5.\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003ereaction setup:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eIt is recommended to assemble the\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003ereaction in a clean bench, and add templates—including cell lysate samples and internal controls—in separate, isolated areas to avoid cross-contamination and aerosol contamination. All experiments should use nuclease-free ultrapure water, pipette tips, and reaction tubes. It is advisable to use filter-tipped pipette tips, especially when adding templates, to prevent cross-contamination from the pipette tip.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eIt is recommended that RT-qPCR be performed in a\u003c\/span\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003ereaction system, assembled on ice. The RT-qPCR Mix reagent in the kit is 5x concentrated; according to the table below, add gene-specific primers (final concentration\u003c\/span\u003e\u003cspan\u003e100-400 nM\u003c\/span\u003e\u003cspan\u003e), and the cell lysis product (\u003c\/span\u003e\u003cspan\u003e0.5-2\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003e, prepared in steps 1-4) and ROX. The amount of ROX should be determined based on the qPCR instrument: for instruments that do not require ROX, add ROX at a high concentration (100x;\u003c\/span\u003e\u003cspan\u003e0.2\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el \/20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003ereaction) or, for instruments requiring low-concentration ROX, add it at 1000x (\u003c\/span\u003e\u003cspan\u003e0.02\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el \/20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003ereaction). For details on major qPCR instruments and ROX concentrations, please refer to the appendix or consult the qPCR instrument's user manual and manufacturer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e20\u003c\/span\u003e\u003cspan\u003eμl\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eRT-qPCR reaction system\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eRT-qPCR Mix (5x)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e4 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eGene-specific primer pair\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIt is recommended that the final concentration of each primer be\u003c\/span\u003e\u003cspan\u003e200 nM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eCell lysate (template)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIt is recommended\u003c\/span\u003e\u003cspan\u003e0.5–1 μl\u003c\/span\u003e\u003cspan\u003e, not exceeding\u003c\/span\u003e\u003cspan\u003e2 μl\u003c\/span\u003e\u003cspan\u003e, with the number of cells not exceeding\u003c\/span\u003e\u003cspan\u003e400\u003c\/span\u003e\u003cspan\u003eindividuals\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eROX\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0 μl\u003c\/span\u003e\u003cspan\u003e(no ROX),\u003c\/span\u003e\u003cspan\u003e0.2 μl\u003c\/span\u003e\u003cspan\u003e(High ROX) or\u003c\/span\u003e\u003cspan\u003e0.02 μl\u003c\/span\u003e\u003cspan\u003e(Low ROX)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eUltra-pure water (nuclease-free)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd to\u003c\/span\u003e\u003cspan\u003e20 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\u003cp style=\"text-align:left\"\u003e\u003cspan\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6.\u003c\/span\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003eAmplification conditions:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eRT-qPCR\u003c\/span\u003e\u003cspan\u003eAmplification is divided into reverse transcription,\u003c\/span\u003e\u003cspan\u003eTaq\u003c\/span\u003e\u003cspan\u003eenzyme activation, and\u003c\/span\u003e\u003cspan\u003ePCR\u003c\/span\u003e\u003cspan\u003eamplification—three stages. It is recommended to perform\u003c\/span\u003e\u003cspan\u003ePCR\u003c\/span\u003e\u003cspan\u003emelting curve analysis after amplification, especially during the optimization of experimental conditions, in order to distinguish between specific and non-specific amplification as well as primer dimers. After reverse transcription,\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eactivate\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eTaq\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eenzyme strictly according to\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e95\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e℃\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e5\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eminutes\u003c\/span\u003e\u003cspan\u003ewill be carried out. The following example\u003c\/span\u003e\u003cspan\u003ePCR\u003c\/span\u003e\u003cspan\u003eamplification conditions are a two-step method; specific conditions need to be optimized based on the template and primers, particularly the annealing\u003c\/span\u003e\u003cspan\u003e\/\u003c\/span\u003e\u003cspan\u003eextension conditions, or amplification can be performed using a\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003estep method.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003eReverse transcription\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003e, 5\u003c\/span\u003e\u003cspan\u003eminutes\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003eTaq\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eenzyme activation\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e95\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003eminutes (a mandatory condition—do not change)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003ePCR\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eamplification\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e40\u003c\/span\u003e\u003cspan\u003ecycles (\u003c\/span\u003e\u003cspan\u003e95\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003e10 sec\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003e60\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003e20sec\/\u003c\/span\u003e\u003cspan\u003eSignal collection)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003eMelting curve\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e1. It is not recommended to alter the amounts of reaction reagents without rigorous validation.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"100T","offer_id":41678336589899,"sku":"UA079018-100T","price":225.0,"currency_code":"USD","in_stock":true},{"title":"1000T","offer_id":41678336622667,"sku":"UA079018-1000T","price":1165.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/761f8c1cdb9f49888790342f622a63f8.png?v=1789110017"},{"product_id":"n-sugar-analysis-kit","title":"N-Glycan Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eBefore starting this protocol, users need to bring the following reagents and equipment (taking 1 IgG sample as an example):\u003cbr\u003e\u003c\/strong\u003e(1) 100uL of 0.85% sodium chloride solution; Acetonitrile (LCMS grade); Ultrapure water;\u003cbr\u003e(2) 15\/85 = ultrapure water\/acetonitrile solution (v\/v), 2 mL; 1\/9\/90 = formic acid\/ultrapure water\/acetonitrile (v\/v), 20 mL; 50 mM ammonium formate, pH = 4.4, 500 mL;\u003cbr\u003e(3) SPE vacuum pump; A 96-well plate negative pressure device or positive pressure device; Heating module or metal bath (90 °C and 50 °C); Vortex instrument; A pipette gun; Centrifuge; Reaction tube.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eStep 1: Rapid Glycosyl Release\u003cbr\u003e\u003c\/strong\u003e(1) The IgG standard and the surfactant in the kit were taken out, and the powder adhered to the tube wall was centrifuged to the bottom of the tube using a centrifuge at 5000 rpm for 15 s.  \u003cbr\u003e(2) Preparation standard: IgG standard (1.2 mg\/5 mg) was dissolved in (600 uL\/2500 uL) 0.85% sodium chloride solution so that the final concentration of IgG standard was 2 mg\/mL.\u003cstrong\u003eNote:\u003c\/strong\u003eUse 0.85% sodium chloride solution or ultrapure water to dissolve the IgG standard.\u003cbr\u003e(3) Preparation of glycosyl release buffer: Dissolve one bottle of surfactant (1.2 mg\/3. 6mg) in (24uL\/72uL) PNGase F buffer, mix well and place at room temperature. (The Glycosyl Release Buffer Solution is Prepared for Ready Use)\u003cbr\u003e(4) Add 20uL of the pre-prepared sample to be tested (2mg\/mL) and IgG standard (2mg\/mL) to the reaction tube, add 3uL of glycosyl release buffer, pipette and mix well for 5 times, then add 3.3 uL of ultrapure water, pipette and mix well for 5 times to mix well.\u003cstrong\u003eNote:\u003c\/strong\u003eThe concentration of the sample to be tested needs to be diluted to 2mg\/mL first, and loading 20uL means the load volume is 40ug. A 200uL reaction tube can be used when heating with a PCR instrument, and a 0.6 mL\/1. 5mL reaction tube can be used according to the instrument specifications when using other heating instruments.\u003cbr\u003e(5) The mixture was incubated at 90 °C for 3 minutes to denature, and after the reaction was completed, the mixture was taken out and allowed to cool at room temperature for 3 minutes.\u003cstrong\u003eNote:\u003c\/strong\u003eThis step requires room temperature cooling for 3 minutes to avoid the excessive temperature of the reaction system affecting the activity of glycosidase in the next step, and the extension of room temperature cooling time will not affect the experiment.\u003cbr\u003e(6) Add another 1.2 uL of PNGase F, pipette and mix well 5 times to mix well, and incubate at 50 °C for 5 minutes.\u003cstrong\u003eNote:\u003c\/strong\u003eGlycosidase (PNGase F) was removed from-20 ° when added to the system. It should not be removed in advance and placed at room temperature to prevent the enzyme activity from decreasing.\u003cbr\u003e(7) After the reaction is completed, take it out and let it cool at room temperature for 3 minutes.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eStep 2: Quickly label glycosyl groups\u003cbr\u003e\u003c\/strong\u003e(1) Labeling reagent: Dissolve 1 bottle of labeling substance (8.2 mg\/24. 6 mg) in (60.13 uL\/180. 4 uL) anhydrous DMF. The labeling reagent is ready for use, and the solution is aspirated and dispersed 5-10 times to completely dissolve. (This mark is currently used and prepared)\u003cstrong\u003eNote:\u003c\/strong\u003eBecause the labeling reagent is placed in a brown bottle, the brown bottle is inconvenient to centrifuge. After adding anhydrous DMF, try to fully dissolve the powder on the upper part of the brown bottle when sucking and mixing.\u003cbr\u003e(2) Add 6uL of labeling reagent solution to the reaction tube, pipette and mix well for 5 times, and react at room temperature for 5 minutes.\u003cbr\u003e(3) Add 179uL of acetonitrile to the reaction tube, pipette and mix well for 5 times, and then use for purification.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eStep 3: Purification\u003cbr\u003e\u003c\/strong\u003e(1) Install negative pressure or positive pressure device and 96-well extraction plate.\u003cstrong\u003eNote:\u003c\/strong\u003eWhen using the vacuum device, the pressure should not be too large, and the appropriate pressure can be set according to different instrument models.\u003cbr\u003e(2) Activation: Add 200uL of ultrapure water to activate the hole to be used, vacuum suction the liquid and collect the waste liquid tray.\u003cbr\u003e(3) Equilibrium: Add 200uL 15\/85 water\/acetonitrile solution to the equilibrium hole, vacuum suction the liquid and collect the waste liquid tray.\u003cbr\u003e(4) Loading: Load the sample diluted with acetonitrile (212.5 uL), and collect the waste liquid tray after vacuum suction.\u003cbr\u003e(5) Add 600uL 1\/9\/90 formic acid\/ultrapure water\/acetonitrile to clean the hole, and collect the waste liquid tray after vacuum suction.\u003cbr\u003e(6) Add 600uL 1\/9\/90 formic acid\/ultrapure water\/acetonitrile to clean the hole, and collect the waste liquid tray after vacuum suction. (i.e., washing twice)\u003cbr\u003e(7) Negative pressure air suction once to suck the residual liquid at the filter element cleanly.\u003cbr\u003e(8) Remove the waste liquid tray and replace it with a collection plate. 50 uL of elution buffer was added to the wells, and the glycosyl groups were eluted by vacuum aspiration to the collection plate, and 50 uL of elution buffer was added to the wells, and the glycosyl groups were eluted by vacuum aspiration to the collection plate. (i.e. final elution volume 100uL)\u003cstrong\u003eNote:\u003c\/strong\u003eThe liquid added to the wells is added to the middle of the wells as much as possible, so as to completely cover the packing and improve the purification efficiency and elution efficiency.\u003cbr\u003e(9) Add 200uL of sample diluent, suck 5 times and mix well, and wait for testing on the machine.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eStep 4: HILIC-FLR detection and analysis\u003cbr\u003e\u003c\/strong\u003e(1) Column ACQUITY UPLC ® Glycan BEH Amide, 130 Å, 1.7 um, 2.1 x 150 mm (waters part number 186004742).\u003cbr\u003e(2) Column temperature: 60 ° C.\u003cbr\u003e(3) Mobile phase A: 50 mM ammonium formate (LC-MS grade recommended) solution, pH = 4.4.\u003cbr\u003e(4) Mobile phase B: 100% acetonitrile (LC-MS grade is recommended).\u003cbr\u003e(5) Flow rate: 0.4 mL\/min.\u003cbr\u003e(6) Gradient:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 79.012%; height: 186px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 44px;\"\u003e\n\u003ctd style=\"width: 19.2384%; text-align: center; height: 44px;\"\u003eTime (min)\u003c\/td\u003e\n\u003ctd style=\"width: 26.4914%; text-align: center; height: 44px;\"\u003eFlow Rate (mL\/min)\u003c\/td\u003e\n\u003ctd style=\"width: 15.6259%; text-align: center; height: 44px;\"\u003e% A\u003c\/td\u003e\n\u003ctd style=\"width: 15.4575%; text-align: center; height: 44px;\"\u003e% B\u003c\/td\u003e\n\u003ctd style=\"width: 14.0727%; text-align: center; height: 44px;\"\u003eCurve\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 19.2384%; height: 22px; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 26.4914%; height: 22px; text-align: center;\"\u003e0.4\u003c\/td\u003e\n\u003ctd style=\"width: 15.6259%; height: 22px; text-align: center;\"\u003e25\u003c\/td\u003e\n\u003ctd style=\"width: 15.4575%; height: 22px; text-align: center;\"\u003e75\u003c\/td\u003e\n\u003ctd style=\"width: 14.0727%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 19.2384%; height: 22px; text-align: center;\"\u003e35\u003c\/td\u003e\n\u003ctd style=\"width: 26.4914%; height: 22px; text-align: center;\"\u003e0.4\u003c\/td\u003e\n\u003ctd style=\"width: 15.6259%; height: 22px; text-align: center;\"\u003e46\u003c\/td\u003e\n\u003ctd style=\"width: 15.4575%; height: 22px; text-align: center;\"\u003e54\u003c\/td\u003e\n\u003ctd style=\"width: 14.0727%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 19.2384%; height: 22px; text-align: center;\"\u003e36.5\u003c\/td\u003e\n\u003ctd style=\"width: 26.4914%; height: 22px; text-align: center;\"\u003e0.2\u003c\/td\u003e\n\u003ctd style=\"width: 15.6259%; height: 22px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 15.4575%; height: 22px; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 14.0727%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 19.2384%; height: 22px; text-align: center;\"\u003e39.5\u003c\/td\u003e\n\u003ctd style=\"width: 26.4914%; height: 22px; text-align: center;\"\u003e0.2\u003c\/td\u003e\n\u003ctd style=\"width: 15.6259%; height: 22px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 15.4575%; height: 22px; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 14.0727%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 19.2384%; height: 22px; text-align: center;\"\u003e43.1\u003c\/td\u003e\n\u003ctd style=\"width: 26.4914%; height: 22px; text-align: center;\"\u003e0.2\u003c\/td\u003e\n\u003ctd style=\"width: 15.6259%; height: 22px; text-align: center;\"\u003e25\u003c\/td\u003e\n\u003ctd style=\"width: 15.4575%; height: 22px; text-align: center;\"\u003e75\u003c\/td\u003e\n\u003ctd style=\"width: 14.0727%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 19.2384%; height: 22px; text-align: center;\"\u003e47.6\u003c\/td\u003e\n\u003ctd style=\"width: 26.4914%; height: 22px; text-align: center;\"\u003e0.4\u003c\/td\u003e\n\u003ctd style=\"width: 15.6259%; height: 22px; text-align: center;\"\u003e25\u003c\/td\u003e\u0026lt; td style = \"width: 15.4575%; height: 22px; text-align: center; \"\u0026gt;75\u003ctd style=\"width: 14.0727%; height: 22px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 19.2384%; height: 10px; text-align: center;\"\u003e55\u003c\/td\u003e\n\u003ctd style=\"width: 26.4914%; height: 10px; text-align: center;\"\u003e0.4\u003c\/td\u003e\n\u003ctd style=\"width: 15.6259%; height: 10px; text-align: center;\"\u003e25\u003c\/td\u003e\n\u003ctd style=\"width: 15.4575%; height: 10px; text-align: center;\"\u003e75\u003c\/td\u003e\n\u003ctd style=\"width: 14.0727%; height: 10px; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e(7) FLR wavelength: EX 265\/EM 425 nm.\u003cbr\u003e(8) FLR sampling rate: 2 Hz.\u003cbr\u003e(9) Injection volume: 10uL.\u003cbr\u003eRemarks: The above parameters are based on the LCMS equipped with \"ACQUITY ® RDa\" detector, and the user adjusts the appropriate parameters according to the equipment.\u003cbr\u003e\u003cbr\u003e\u003cbr\u003e\u003cbr\u003e\u003cbr\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eAbsin provides information on the routine maintenance and use of N-sugar analysis kits for rapid enzymatic release and rapid labeling of N-sugars. This protocol has been validated using monoclonal antibodies and has been tested for use with a variety of other N-linked glycoproteins. We recommend that users confirm the enzyme release of their own specific samples.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eProduct Components:\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 82.0899%; height: 294px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 19.8681%; text-align: center; height: 21px;\"\u003e\u003cstrong\u003eModule\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 38.6362%; text-align: center; height: 21px;\"\u003e\u003cstrong\u003eComponents\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; text-align: center; height: 21px;\"\u003e\u003cstrong\u003e24T\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; text-align: center; height: 21px;\"\u003e\u003cstrong\u003e96 T\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"height: 84px; width: 19.8681%; text-align: center;\" rowspan=\"4\"\u003eGlycosyl release module\u003c\/td\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003eIgG\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e1.2 mg\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e5mg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003ePNGase F enzyme\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e45uL\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e150uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003ePNGase F Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e0.5 mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003eSurfactant\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e1.2 mg × 6\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e3.6 mg × 8\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"height: 42px; width: 19.8681%; text-align: center;\" rowspan=\"2\"\u003eTag module\u003c\/td\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003eMarker\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e8.2 mg × 3\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e24.6 mg × 4\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003eAnhydrous DMF\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"height: 21px; width: 19.8681%; text-align: center;\" rowspan=\"3\"\u003ePurification module\u003c\/td\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003e96-well extraction plate\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e1 piece\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e1 piece\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003eElution Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e5 mL × 1\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e5 mL × 4\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003eSample dilution\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e5 mL × 1\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e5 mL × 4\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"height: 21px; width: 19.8681%; text-align: center;\" rowspan=\"4\"\u003eCollection module\u003c\/td\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003e8-row reaction tubes (200uL)\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003e8 connected cover\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e100\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003eCollection plate (96 well)\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 38.6362%; height: 21px; text-align: center;\"\u003eWaste tray\u003c\/td\u003e\n\u003ctd style=\"width: 17.839%; height: 21px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 16.3395%; height: 21px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. For your safety and health, please wear a laboratory coat and gloves.\u003cbr\u003e2. Please use a clean pipette tip every time you take it.\u003cbr\u003e3. Please store and use relevant reagents according to the instructions.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 82.669%; height: 240px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; text-align: center; height: 21px;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; text-align: center; height: 21px;\"\u003ePreservation conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 21px; text-align: center;\"\u003eIgG\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 21px; text-align: center;\"\u003e2-8 °C, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 21px; text-align: center;\"\u003ePNGase F\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 21px; text-align: center;\"\u003e-25 °C--15 °C, 12 months, avoid repeated freezing and thawing\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 21px; text-align: center;\"\u003ePNGase F Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 21px; text-align: center;\"\u003e2-8 °C, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 21px; text-align: center;\"\u003eSurfactant\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 21px; text-align: center;\"\u003eAmbient temperature, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 21px; text-align: center;\"\u003eMarker\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 21px; text-align: center;\"\u003e2-8 °C, 12 months, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 21px; text-align: center;\"\u003eAnhydrous DMF\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 21px; text-align: center;\"\u003eAmbient temperature, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 21px; text-align: center;\"\u003eElution Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 21px; text-align: center;\"\u003eAmbient temperature, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 21px; text-align: center;\"\u003eSample dilution\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 21px; text-align: center;\"\u003eAmbient temperature, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 10px; text-align: center;\"\u003e96-well extraction plate\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 10px; text-align: center;\"\u003eNormal temperature. Once opened, unused holes are sealed with sealing film and placed in a dry environment\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 41px;\"\u003e\n\u003ctd style=\"width: 35.5487%; height: 41px; text-align: center;\"\u003e200uL 8-row reaction tube, 8-row cover, 96-well collection plate, waste liquid tray\u003c\/td\u003e\n\u003ctd style=\"width: 60.8082%; height: 41px; text-align: center;\"\u003eAt room temperature until the end of the validity period, 96-well collection plate is sealed and stored dry.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"24T","offer_id":41683172819019,"sku":"abs590006-24T","price":2133.0,"currency_code":"USD","in_stock":true},{"title":"96T","offer_id":41683172851787,"sku":"abs590006-96T","price":7133.0,"currency_code":"USD","in_stock":true},{"title":"5T","offer_id":41683172884555,"sku":"abs590006-5T","price":550.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/bc3daf446ce24170aebe260c86d408d8.jpg?v=1789743623"},{"product_id":"n-sugar-analysis-kit-abs590007","title":"N-Glycan Kit(Extraction column)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003eBefore starting this protocol, users need to bring the following reagents and equipment (taking an IgG sample as an example):\u003c\/strong\u003e\u003cbr\u003e(1) 100uL of 0.85% sodium chloride solution; Acetonitrile (LCMS grade); Ultrapure water;  \u003cbr\u003e(2) 15\/85 = ultrapure water\/acetonitrile solution (v\/v), 2 mL; 1\/9\/90 = formic acid\/ultrapure water\/acetonitrile (v\/v), 20 mL; 50 mM ammonium formate, PH = 4.4, 500 mL;  \u003cbr\u003e(3) SPE vacuum pump; A solid phase extraction device; Heating module or metal bath (90 °C and 50 °C); Vortex instrument; A pipette gun; Centrifuge; Reaction tube.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eStep 1: Rapid Glycosyl Release\u003cbr\u003e\u003c\/strong\u003e(1) The IgG standard and the surfactant in the kit were taken out, and the powder adhered to the tube wall was centrifuged to the bottom of the tube using a centrifuge at 5000 rpm for 15 s.\u003cbr\u003e(2) Preparation standard: IgG standard (0.25 mg\/1. 2 mg) was dissolved in (125 uL\/600 uL) 0.85% sodium chloride solution so that the final concentration of IgG standard was 2 mg\/mL.\u003cstrong\u003eNote:\u003c\/strong\u003eUse 0.85% sodium chloride solution or ultrapure water to dissolve the IgG standard.\u003cbr\u003e(3) Preparation of glycosyl release buffer: Dissolve one bottle of surfactant (1.5 mg\/1. 2mg) in (30uL\/24uL) PNGase F buffer, mix well and place at room temperature. (The Glycosyl Release Buffer Solution is Prepared for Ready Use)\u003cbr\u003e(4) Add 20uL of the pre-prepared sample to be tested (2mg\/mL) and IgG standard (2mg\/mL) to the reaction tube, add 3uL of glycosyl release buffer, pipette and mix well for 5 times, then add 3.3 uL of ultrapure water, pipette and mix well for 5 times to mix well.\u003cstrong\u003eNote:\u003c\/strong\u003eThe concentration of the sample to be tested needs to be diluted to 2mg\/mL first, and loading 20uL means the load volume is 40ug. A 200uL reaction tube can be used when heating with a PCR instrument, and a 0.6 mL\/1. 5mL reaction tube can be used according to the instrument specifications when using other heating instruments.\u003cbr\u003e(5) The mixture was incubated at 90 °C for 3 minutes to denature, and after the reaction was completed, the mixture was taken out and allowed to cool at room temperature for 3 minutes.\u003cstrong\u003eNote:\u003c\/strong\u003eThis step requires room temperature cooling for 3 minutes to avoid the excessive temperature of the reaction system affecting the activity of glycosidase in the next step, and the extension of room temperature cooling time will not affect the experiment.\u003cbr\u003e(6) Add another 1.2 uL of PNGase F, pipette and mix well 5 times to mix well, and incubate at 50 °C for 5 minutes.\u003cstrong\u003eNote:\u003c\/strong\u003eGlycosidase (PNGase F) was removed from-20 ° when added to the system. It should not be removed in advance and placed at room temperature to prevent the enzyme activity from decreasing.\u003cbr\u003e(7) After the reaction is completed, take it out and let it cool at room temperature for 3 minutes.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eStep 2: Quickly label glycosyl groups\u003cbr\u003e\u003c\/strong\u003e(1) Configuration of labeling reagent: Dissolve 1 bottle of labeling substance (4.77 mg\/8. 2mg) in (35.10 uL\/60. 13uL) anhydrous DMF, pipping and mixing 5-10 times for complete dissolution. (The labeling reagent is now prepared and used)\u003cstrong\u003eNote:\u003c\/strong\u003eBecause the labeling reagent is placed in a brown bottle, the brown bottle is inconvenient to centrifuge. After adding anhydrous DMF, try to fully dissolve the powder on the upper part of the brown bottle when sucking and mixing.\u003cbr\u003e(2) Add 6uL of labeling reagent to the reaction tube, pipette and mix well for 5 times, and react at room temperature for 5 minutes.\u003cbr\u003e(3) Add 179uL of acetonitrile to the reaction tube, pipette and mix well for 5 times, and then use for purification.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eStep 3: Purification\u003c\/strong\u003e\u003cbr\u003e(1) Take out the solid phase extraction cartridge in the kit and place it on the fixed frame.\u003cbr\u003e(2) Activation: Add 200μL of ultrapure water to activate the hole to be used, blow the liquid with a pipette gun or ear cleaning ball, and collect the waste liquid tray.\u003cstrong\u003eNote:\u003c\/strong\u003eWhen using a pipette gun or ear washing ball, you need to pay attention: first press the interface between the pipette gun\/ear washing ball and the extraction cartridge tightly, and then blow the liquid vigorously; After blowing, first loosen the interface between the pipette\/ear washing ball and the extraction cartridge, and then loosen the pipette\/ear washing ball. Avoid the collapse of the packing due to the air retraction of the pipette gun\/ear cleaning ball, which will affect the purification of the sample.\u003cbr\u003e(3) Equilibrium: Add 200uL 15\/85 water\/acetonitrile solution to the equilibrium hole, blow the liquid with a pipette gun or ear cleaning ball and collect the waste liquid tray.\u003cbr\u003e(4) Loading: Load the sample diluted with acetonitrile (212.5 uL), blow the liquid with a pipette gun or ear cleaning ball and collect it in the waste tray.\u003cbr\u003e(5) Add 600uL 1\/9\/90 formic acid\/ultrapure water\/acetonitrile to clean the hole, blow the liquid with a pipette gun or ear cleaning ball and collect the waste liquid tray.\u003cbr\u003e(6) Add 600uL 1\/9\/90 formic acid\/ultrapure water\/acetonitrile to clean the hole, blow the liquid with a pipette gun or ear cleaning ball and collect the waste liquid tray. (i.e., washing twice)\u003cbr\u003e(7) Blow the pipette gun or ear cleaning ball once to clean the residual liquid at the filter element.\u003cbr\u003e(8) Remove the waste liquid tray and replace it with a 1.5 mL reaction tube, add 50uL of elution buffer into the hole, collect the liquid after pipetting the liquid with a pipette gun or ear washing ball, and then add 50uL of elution buffer into the hole, collect the liquid after pipetting the liquid with a pipette gun or ear washing ball. (i.e. final elution volume 100uL)\u003cstrong\u003eNote:\u003c\/strong\u003eThe liquid added to the wells is added to the middle of the wells as much as possible, so as to completely cover the packing and improve the purification efficiency and elution efficiency.\u003cbr\u003e(9) Add 100uL of sample diluent, suck 5 times and mix well, and wait for testing on the machine.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eStep 4: HILIC-FLR detection and analysis\u003c\/strong\u003e\u003cbr\u003e(1) Column ACQUITY UPLC ® Glycan BEH Amide, 130 Å, 1.7 um, 2.1 x 150 mm (waters part number 186004742).\u003cbr\u003e(2) Column temperature: 60 ° C.\u003cbr\u003e(3) Mobile phase A: 50 mM ammonium formate (LC-MS grade recommended) solution, pH = 4.4.\u003cbr\u003e(4) Mobile phase B: 100% acetonitrile (LC-MS grade is recommended).\u003cbr\u003e(5) Flow rate: 0.4 mL\/min.\u003cbr\u003e(6) Gradient:\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 57.1056%; height: 202px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 45px;\"\u003e\n\u003ctd style=\"width: 18.2329%; text-align: center; height: 45px;\"\u003eTime (min)\u003c\/td\u003e\n\u003ctd style=\"width: 16.6371%; text-align: center; height: 45px;\"\u003eFlow Rate (mL\/min)\u003c\/td\u003e\n\u003ctd style=\"width: 14.1229%; text-align: center; height: 45px;\"\u003e% A\u003c\/td\u003e\n\u003ctd style=\"width: 14.0877%; text-align: center; height: 45px;\"\u003e% B\u003c\/td\u003e\n\u003ctd style=\"width: 17.6084%; text-align: center; height: 45px;\"\u003eCurve\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 23px;\"\u003e\n\u003ctd style=\"width: 18.2329%; height: 23px;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 16.6371%; height: 23px;\"\u003e0.4\u003c\/td\u003e\n\u003ctd style=\"width: 14.1229%; height: 23px;\"\u003e25\u003c\/td\u003e\n\u003ctd style=\"width: 14.0877%; height: 23px;\"\u003e75\u003c\/td\u003e\n\u003ctd style=\"width: 17.6084%; height: 23px;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 23px;\"\u003e\n\u003ctd style=\"width: 18.2329%; height: 23px;\"\u003e35\u003c\/td\u003e\n\u003ctd style=\"width: 16.6371%; height: 23px;\"\u003e0.4\u003c\/td\u003e\n\u003ctd style=\"width: 14.1229%; height: 23px;\"\u003e46\u003c\/td\u003e\n\u003ctd style=\"width: 14.0877%; height: 23px;\"\u003e54\u003c\/td\u003e\n\u003ctd style=\"width: 17.6084%; height: 23px;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 23px;\"\u003e\n\u003ctd style=\"width: 18.2329%; height: 23px;\"\u003e36.5\u003c\/td\u003e\n\u003ctd style=\"width: 16.6371%; height: 23px;\"\u003e0.2\u003c\/td\u003e\n\u003ctd style=\"width: 14.1229%; height: 23px;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 14.0877%; height: 23px;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 17.6084%; height: 23px;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 23px;\"\u003e\n\u003ctd style=\"width: 18.2329%; height: 23px;\"\u003e39.5\u003c\/td\u003e\n\u003ctd style=\"width: 16.6371%; height: 23px;\"\u003e0.2\u003c\/td\u003e\n\u003ctd style=\"width: 14.1229%; height: 23px;\"\u003e100\u003c\/td\u003e\n\u003ctd style=\"width: 14.0877%; height: 23px;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 17.6084%; height: 23px;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 23px;\"\u003e\n\u003ctd style=\"width: 18.2329%; height: 23px;\"\u003e43.1\u003c\/td\u003e\n\u003ctd style=\"width: 16.6371%; height: 23px;\"\u003e0.2\u003c\/td\u003e\n\u003ctd style=\"width: 14.1229%; height: 23px;\"\u003e25\u003c\/td\u003e\n\u003ctd style=\"width: 14.0877%; height: 23px;\"\u003e75\u003c\/td\u003e\n\u003ctd style=\"width: 17.6084%; height: 23px;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 24px;\"\u003e\n\u003ctd style=\"width: 18.2329%; height: 24px;\"\u003e47.6\u003c\/td\u003e\n\u003ctd style=\"width: 16.6371%; height: 24px;\"\u003e0.4\u003c\/td\u003e\n\u003ctd style=\"width: 14.1229%; height: 24px;\"\u003e25\u003c\/td\u003e\n\u003ctd style=\"width: 14.0877%; height: 24px;\"\u003e75\u003c\/td\u003e\n\u003ctd style=\"width: 17.6084%; height: 24px;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 18.2329%; height: 18px;\"\u003e55\u003c\/td\u003e\n\u003ctd style=\"width: 16.6371%; height: 18px;\"\u003e0.4\u003c\/td\u003e\n\u003ctd style=\"width: 14.1229%; height: 18px;\"\u003e25\u003c\/td\u003e\n\u003ctd style=\"width: 14.0877%; height: 18px;\"\u003e75\u003c\/td\u003e\n\u003ctd style=\"width: 17.6084%; height: 18px;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e(7) FLR wavelength: EX 265\/EM 425 nm.\u003cbr\u003e(8) FLR sampling rate: 2 Hz.\u003cbr\u003e(9) Injection volume: 10uL. \u003cbr\u003e\u003cbr\u003eRemarks: The above parameters are based on LCMS equipped with \"ACQUITY ® RDa\" detector, and the user adjusts the appropriate parameters according to the equipment.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eABISIN provides information on the routine maintenance and use of N-sugar analysis kits for rapid enzymatic release and rapid labeling of N-sugars. This protocol has been validated using monoclonal antibodies and has been tested for use with a variety of other N-linked glycoproteins. We recommend that users confirm the enzyme release of their own specific samples.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eProduct Nature:\u003cbr\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 60.0603%; height: 270px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 26.7464%; height: 21px;\"\u003e\u003cstrong\u003eModule\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 40.041%; height: 21px;\"\u003e\u003cstrong\u003eComponents\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 21px;\"\u003e\u003cstrong\u003e24T\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 26.7464%; height: 85px;\" rowspan=\"4\"\u003eGlycosyl release module\u003c\/td\u003e\n\u003ctd style=\"width: 40.041%; height: 21px;\"\u003eIgG\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 21px;\"\u003e1.2 mg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 40.041%; height: 21px;\"\u003ePNGase F enzyme\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 21px;\"\u003e45uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 40.041%; height: 21px;\"\u003ePNGase F Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 21px;\"\u003e0.5 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 40.041%; height: 22px;\"\u003eSurfactant\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 22px;\"\u003e1.2 mg × 6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 26.7464%; height: 44px;\" rowspan=\"2\"\u003eTag module\u003c\/td\u003e\n\u003ctd style=\"width: 40.041%; height: 22px;\"\u003eMarker\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 22px;\"\u003e8.2 mg × 3\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 40.041%; height: 22px;\"\u003eAnhydrous DMF\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 22px;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 26.7464%; height: 66px;\" rowspan=\"3\"\u003ePurification module\u003c\/td\u003e\n\u003ctd style=\"width: 40.041%; height: 22px;\"\u003eSolid phase extraction cartridge\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 22px;\"\u003e8 pieces\/pack × 3\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 40.041%; height: 22px;\"\u003eElution Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 22px;\"\u003e5 mL × 1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 40.041%; height: 22px;\"\u003eSample dilution\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 22px;\"\u003e5 mL × 1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 26.7464%; height: 54px;\" rowspan=\"3\"\u003eCollection module\u003c\/td\u003e\n\u003ctd style=\"width: 40.041%; height: 22px;\"\u003e8-row reaction tubes (200uL)\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 22px;\"\u003e100\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 40.041%; height: 22px;\"\u003e8 connected cover\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 22px;\"\u003e100\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px;\"\u003e\n\u003ctd style=\"width: 40.041%; height: 10px;\"\u003eWaste tray\u003c\/td\u003e\n\u003ctd style=\"width: 23.6327%; height: 10px;\"\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cstrong\u003e \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. For your safety and health, please wear a laboratory coat and gloves.\u003cbr\u003e2. Please use a clean pipette tip every time you take it.\u003cbr\u003e3. Please store and use relevant reagents according to the instructions.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003ctable style=\"border-collapse: collapse; width: 60.7562%; height: 412px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%; text-align: center;\"\u003ePreservation conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003eIgG\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003e2-8 °C, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003ePNGase F\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003e-25 °C--15 °C, 12 months, avoid repeated freezing and thawing\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003ePNGase F Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003e2-8 °C, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003eSurfactant\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003eAmbient temperature, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003eMarker\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003e2-8 °C, 12 months, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003eAnhydrous DMF\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003eAmbient temperature, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003eElution Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003eAmbient temperature, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003eSample dilution\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003eAmbient temperature, 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003eSolid phase extraction cartridge\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003eAt room temperature, the unused cartridge is sealed and stored\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 42.3139%;\"\u003e200uL 8-row reaction tube, 8-row cover, 96-well collection plate, waste liquid tray\u003c\/td\u003e\n\u003ctd style=\"width: 51.0898%;\"\u003eNormal temperature to the end of validity\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"24T","offer_id":41683172950091,"sku":"abs590007-24T","price":2133.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/d96491f14a234c5b950e595c80822aef.jpg?v=1789743624"},{"product_id":"tissue-digestion-kit-ua070090","title":"Tissue Digestion Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePurity\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e≥95% by SDS-PAGE\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eEndotoxin\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u0026lt;10 EU\/mg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:12pt;color:rgb(0,0,0)\"\u003e\u003c\/span\u003e\u003cspan style=\"color: rgb(0, 0, 0); font-size: 14px;\"\u003eThis product should be stored at -20℃ and can be stored for at least 12 months.\u003c\/span\u003e\u003cspan style=\"font-size:12pt;color:rgb(0,0,0)\"\u003e\u003cbr\u003e\u003c\/span\u003e\u003cspan style=\"color: rgb(0, 0, 0); font-size: 14px;\"\u003eAvoid repeated freezing and thawing\u003c\/span\u003e\u003cspan style=\"font-size:12pt;color:rgb(0,0,0)\"\u003e\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eThis kit is composed of recombinant collagenase mixed components and recombinant porcine trypsin. The recombinant collagenase mixed components are expressed in E. coli and are specific to the bond between the neutral amino acid (X) and glycine in the collagen-specific amino acid sequence Pro-X-Gly-Pro; the recombinant porcine trypsin is recombinantly expressed in Pichia pastoris and can specifically cleave the carboxyl-terminal peptide bonds of lysine and arginine in proteins. Both the recombinant collagenase mixed components and the recombinant porcine trypsin do not contain animal-derived ingredients, and can be used to efficiently digest animal organs such as the heart, liver, and kidney to obtain corresponding primary cells.\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 684.34px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 247.6px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eComponents\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 138.007px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eS\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 143.009px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eM\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 155px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eL\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.183px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eRecombinant collagenase mixed components\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e1.0ml×2\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e4.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.229px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e10.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.183px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eRecombinant porcine trypsin\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e8.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e16.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.229px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e40.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 676.34px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 134.6px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eSize\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 236.005px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eCollagenase activity\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 305.009px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eTrypsin activity\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eS\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;3500CDU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;25KU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eM\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;\u003c\/span\u003e\u003c\/font\u003e\u003cspan style=\"font-size: 14px;\"\u003e7000CDU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;\u003c\/span\u003e\u003c\/font\u003e\u003cspan style=\"font-size: 14px;\"\u003e50KU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eL\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;\u003c\/span\u003e\u003c\/font\u003e\u003cspan style=\"font-size: 14px;\"\u003e17500CDU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;\u003c\/span\u003e\u003c\/font\u003e\u003cspan style=\"font-size: 14px;\"\u003e125KU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eInstructions for use (digestion of mouse heart tissue, for reference only)\u003c\/span\u003e\u003c\/p\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eThaw the recombinant collagenase mixed components and recombinant porcine trypsin in the kit and mix them in a sterile state at a volume ratio of 1:4 to prepare a digestion solution. If further dilution is required, HBSS or PBS buffer can be used for dilution.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRemove the mouse heart from the tissue preservation solution and place it in pre-cooled PBS (+2% blue chain double antibody) and wash 2-3 times.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRemove the excess part of the tissue (such as connective tissue, adipose tissue, etc.).\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eCut the mouse heart into pieces (about 3mm3), transfer it into a centrifuge tube, add 1.5ml of digestion solution, digest at 37℃ for 20 minutes, and blow or invert the pipette every 10 minutes to mix well to fully digest the tissue. During this process, the digestion situation needs to be carefully checked to avoid over-digestion. If necessary, the digestion suspension can be examined under a microscope. It is best to observe more single cells or smaller cell clusters (\u0026lt;70µm) under the microscope.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eLet the digestion suspension stand for a while, collect the supernatant, add 1.5ml digestion solution to the precipitate, digest at 37℃ for 20 minutes, and mix by blowing or inverting with a pipette every 10 minutes or so to fully digest the tissue.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRepeat the above steps once.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eCombine the collected supernatants, add 5ml DMEM complete medium to terminate the digestion, centrifuge at 4℃, 300g for 5 minutes, and discard the supernatant.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eResuspend the cell pellet with 5ml PBS (+1% blue chain double antibody), pass through a 70µm cell sieve, and centrifuge at 4℃, 300g for 5 minutes.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eResuspend the cell pellet with 5ml PBS (+1% blue chain double antibody), centrifuge at 4℃, 300g for 5 minutes, and discard the supernatant.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\n\u003cspan style=\"font-size:10.5000pt\"\u003eResuspend the cell pellet with an appropriate volume of DMEM complete medium and culture it in a cell culture dish.\u003c\/span\u003e\u003cbr\u003e\n\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cspan style=\"font-size: 14px;\"\u003eIf more cell clumps than single cells are required, shortening the digestion time or reducing the proportion of trypsin is recommended;\u003c\/span\u003e\u003cbr\u003e\u003cspan style=\"font-size: 14px;\"\u003eTry to avoid repeated freezing and thawing of this product. If there is any surplus, it is recommended to pack and store at -20℃\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eUnit Definition\u003c\/h4\u003e\u003cdiv\u003eCollagenase activity definition: Within 5 hours, at pH 7.4, 37 °C and in the presence of calcium ions, peptides are released from collagen of bovine Achilles tendon, and the amount of enzyme equivalent to 1.0 µmol of leucine in ninhydrin is 1 CDU.Trypsin activity definition: At 25 °C, pH 7.6, reaction system 3.2 ml (1 cm light path), the amount of enzyme that hydrolyzes BAEE per minute to increase the absorbance at 253 nm by 0.003 is defined as 1 U.\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"S","offer_id":41738013474891,"sku":"UA070090-S","price":295.0,"currency_code":"USD","in_stock":true},{"title":"M","offer_id":41738013507659,"sku":"UA070090-M","price":420.0,"currency_code":"USD","in_stock":true},{"title":"L","offer_id":41738013540427,"sku":"UA070090-L","price":795.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/edc44bd8-21f8-44e8-be9f-7ec93eea0e18.png?v=1787969027"},{"product_id":"tissue-digestion-kit-ii-ua070091","title":"Tissue Digestion Kit II","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePurity\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e≥95% by SDS-PAGE\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eEndotoxin\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u0026lt;10 EU\/mg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:12pt;color:rgb(0,0,0)\"\u003eThis product should be stored at -20℃ and can be stored for at least 12 months.Avoid repeated freezing and thawing\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eThis kit is composed of a recombinant collagenase mixed component and a heat-resistant metalloprotease. The recombinant collagenase mixed component is expressed in E. coli and has specificity for the bond between the neutral amino acid (X) and glycine in the collagen-specific amino acid sequence Pro-X-Gly-Pro; the heat-resistant metalloprotease is recombinantly expressed in E. coli and can hydrolyze the protein bond at the N-terminus of the hydrophobic amino acid residue. Both the recombinant collagenase mixed component and the heat-resistant metalloprotease do not contain animal-derived ingredients. At the same time, they can efficiently digest animal organs such as the intestine, heart, liver, and kidney to obtain corresponding primary cells. Compared with the kit using trypsin, the activity of the heat-resistant metalloprotease is not affected by serum, which is more suitable for scenarios where serum needs to be added during the digestion process.\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 780.34px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eComponents\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 164.417px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eS\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 162.007px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eM\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 155.005px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eL\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.183px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eRecombinant collagenase mixed components\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e1.0ml×2\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e4.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.229px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e10.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.183px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eThermoresistant metalloproteinases\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e1.0ml×6\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.194px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e12.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.229px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e30.0ml\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 754.34px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 128.6px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eSize\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 264px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eCollagenase activity\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 361.007px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eThermostable metalloproteinase activity\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eS\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;3500CDU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;\u003c\/span\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e600U\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eM\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;\u003c\/span\u003e\u003c\/font\u003e\u003cspan style=\"font-size: 14px;\"\u003e7000CDU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;1200U\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.593px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003eL\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;\u003c\/span\u003e\u003c\/font\u003e\u003cspan style=\"font-size: 14px;\"\u003e17500CDU\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.604px;\"\u003e\u003cp\u003e\u003cfont\u003e\u003cspan style=\"font-size: 14px;\"\u003e\u0026gt;3000U\u003c\/span\u003e\u003c\/font\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eInstructions for use (digestion of mouse intestinal tissue, for reference only)\u003c\/span\u003e\u003c\/p\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003ePreparation of digestive solution: 0.5ml recombinant collagenase mixed components + 1.5ml heat-resistant metalloproteinase, dilute with 4.0ml DMEM medium, and add FBS to a final concentration of 3%, DNase I To a final concentration of 100U\/ml, and the blue chain double antibody to a final concentration of 0.1%;\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRemove the small intestine from the tissue preservation solution, transfer it to a 10cm dish, add 10ml pre-cooled PBS + 1% blue chain double antibody, and rinse it 2-3 times;\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eTransfer the rinsed small intestine to a new 10cm dish containing 10ml pre-cooled PBS + 1% blue chain double antibody, rinse it back and forth 3 times with a syringe to remove the contents of the small intestine;\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eTransfer the rinsed small intestine to a new 10cm dish containing 10ml pre-cooled PBS + 1% blue chain double antibody, cut the small intestine longitudinally with scissors, and gently scrape off the villi on the surface of the small intestine with tweezers;\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eTransfer the small intestine to a new 10cm dish containing 10ml pre-cooled PBS + 1% blue chain double antibody, cut the small intestine into 1cm long segments, and transfer it to 2ml EP tube, cut into pieces with scissors appropriately; \u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eTransfer the cut small intestine to a 15ml centrifuge tube, add 6ml digestion solution, and place in a 37℃ water bath for 10min. Take it out after 5min, mix it upside down and let it stand for a while, take the supernatant to a 50ml centrifuge tube, and add DMEM to complete the culture to 20ml; l \u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eAdd 5ml DMEM+3% FBS to the precipitate, blow it several times, let it stand for a while, take the supernatant and combine it with the solution in the previous step; l \u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\n\u003cspan style=\"font-size:10.5000pt\"\u003eCentrifuge at 340g and 4℃ for 8min, discard the supernatant, resuspend it with 12.5ml PBS and pass it through a 70µm cell sieve, and add 8ml DMEM complete culture medium to the sieved cells.\u003c\/span\u003e\u003cbr\u003e\n\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eThis product should avoid repeated freezing and thawing. If there is any surplus, it is recommended to store it at -20℃\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eUnit Definition\u003c\/h4\u003e\u003cdiv\u003eCollagenase activity definition: Within 5 hours, at pH 7.4, 37 °C and in the presence of calcium ions, the amount of enzyme that releases peptides from bovine Achilles tendon collagen and the ninhydrin color is equivalent to 1.0 µmol of leucine is 1 CDU.Thermostable metalloproteinase activity definition: at pH 7.5 and 37 °C, the amount of enzyme required to hydrolyze casein and produce 1.0 μM (181 μg) tyrosine per minute is 1U.\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"S","offer_id":41738013573195,"sku":"UA070091-S","price":415.0,"currency_code":"USD","in_stock":true},{"title":"M","offer_id":41738013605963,"sku":"UA070091-M","price":645.0,"currency_code":"USD","in_stock":true},{"title":"L","offer_id":41738013638731,"sku":"UA070091-L","price":1240.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/ebe73071-dff4-48be-a578-cf402a15eb06.png?v=1787969030"},{"product_id":"startscript-one-step-rt-pcr-kit-ua070126","title":"StartScript® One Step RT-PCR Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 645.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 231.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 209.002px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070126-50 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 204.009px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070126-250 Rxns\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOne-Step Enzyme Mix (25X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e 5 × 100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOne-Step Reaction Mix (2X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 × 625 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 × 625 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 × 1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 × 1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e1. Prepare the following mixture in an RNase-free microcentrifuge tube:  \u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 509.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 319.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 189.005px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOne-Step Enzyme Mix (25X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eOne-Step Reaction Mix (2X)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e25 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eGene-specific Forward Primer (10 μM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eGene-specific Reverse Primer (10 μM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eTemplate RNA (1 pg~1 μg)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003ex μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase-Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 596.435px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUp to 50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e  Mix gently by pipetting.  2. Perform the RT-PCR reaction under the following conditions:  \u003ctable class=\"table table-bordered\" style=\"width: 773.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 225.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eStep\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 143.009px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eTemperature\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 220.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eTime\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 184.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eNumber of Cycles\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eReverse Transcription*\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e48°C\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e30 minutes\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 cycle\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eInitial Denaturation\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e94°C\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e3 minutes\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 cycle\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eDenaturation\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eAnnealing**\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eExtension\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e94°C\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e50-65°C\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e72°C\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e30 seconds\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e30 seconds\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 minute\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e25–35 cycles\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eFinal Extension\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e72°C\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 minutes\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 cycle\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eSoak\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e4°C\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eIndefinite\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 cycle\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e  *If the template has complex secondary structures or high GC content, the reaction temperature can be increased to 55°C to improve yield.  **The annealing temperature should be adjusted according to the primer's melting temperature (Tm), typically set 1–2°C below the primer Tm.\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e1. Treat all equipment used in the study with DEPC or purchase equipment certified to be nucleic acid-free. Wear gloves during the study and change them frequently to avoid RNase contamination.  2. Ensure that the reagents used are free of RNase contamination.  3. Typically, 28–30 cycles achieve optimal amplification. For low-copy target gene detection, the number of cycles can be increased to 40.  4. When performing reverse transcription reactions using this kit, specific reverse transcription primers must be used. Random Primer and Oligo dT Primer cannot be used.  5. Enzymes should be kept on ice during experimental operations and immediately stored at -20°C after the experiment.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50Rxns","offer_id":41738019799115,"sku":"UA070126-50Rxns","price":75.0,"currency_code":"USD","in_stock":true},{"title":"50Rxns*5","offer_id":41738019831883,"sku":"UA070126-50Rxns*5","price":250.0,"currency_code":"USD","in_stock":true},{"title":"1000Rxns","offer_id":43086268170315,"sku":"UA070126-1000Rxns","price":750.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/947f67329785436a83e04490d6b4ba5e.png?v=1788044473"},{"product_id":"biotin-labeling-efficiency-detection-kit-haba-method-ua080468","title":"Biotin Labeling Efficiency Detection Kit (HABA Method)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eBiotin Labeling Efficiency Assay Kit (HABA Method)\u003c\/td\u003e\n\u003c\/tr\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cb\u003e\u003cspan style=\"font-weight: bold;\"\u003e\u003cfont\u003eDetection Principle\u003c\/font\u003e\u003c\/span\u003e\u003c\/b\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cb\u003e\u003cspan style=\"font-weight: bold;\"\u003e   \u003c\/span\u003e\u003c\/b\u003e\u003cspan style=\"font-weight: normal;\"\u003e \u003cfont\u003eDue to the high affinity of biotin for avidin, biotin can displace the interaction between 4'-hydroxyazobenzene-2-carboxylic acid (HABA) and avidin, resulting in a proportional decrease in absorbance at 500 nm. The change in absorbance is correlated with the biotin concentration.\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: normal;\"\u003e    \u003cfont\u003eWhen a biotin-labeled protein solution is added to the HABA-avidin mixture, the change in absorbance before and after the addition of the biotin sample can be measured. According to the Lambert-Beer law, the biotin content in the solution can be calculated.\u003c\/font\u003e\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003cbr\u003eUsed in conjunction with the Rapid Biotin Conjugation Kit to evaluate biotin labeling efficiency.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eAdvantages\u003c\/strong\u003e\u003cbr\u003e(1) Reliable—A mature and validated detection method\u003cbr\u003e(2) Convenient—The kit provides all necessary reagents, including microplates\u003cbr\u003e(3) Flexible—Freedom to choose between spectrophotometry or microplate detection methods\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003eHere is the translated content with original formatting preserved:```html\u003c\/p\u003e\n\u003cp\u003e5T (Spectrophotometry)\u003cbr\u003e24T (Microplate Reader Method)\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 594.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 133.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 156.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eName\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 149.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eVolume\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 155.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eStorage Temperature\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent A\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eHABA Solution\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e200µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent B\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eAvidin\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2.5mg\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003ePhosphate Buffered Saline (PBS)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e10mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eComponent D\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eMicroplate\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1 plate\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eRT\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e```\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eHere is the translated content in English, maintaining the original formatting:---**Unprovided Essentials:**  - Spectrophotometer or microplate reader with 500nm detection capability.  - Pipettes (20-1000µL).  **Reagent Preparation**  1. **Sample**: Before analysis, biotin-labeled protein samples must be desalted or dialyzed to remove all unreacted and hydrolyzed biotin conjugates.  2. **HABA-Avidin Solution Preparation**:     1) Add 480µL of Reagent C (phosphate buffer, PBS) to the tube of Reagent B (avidin) to prepare a 5.2mg\/mL avidin solution.     2) Calculate the required volume of HABA-Avidin mixed solution based on the measurement method (spectrophotometry\/microplate assay): spectrophotometry requires 900µL per test; microplate assay requires 180µL per well. Prepare accordingly.     3) Prepare the HABA-Avidin mixed solution: Mix 138µL of Reagent A (HABA solution) with 460µL of the freshly prepared avidin solution and add to 4002µL of Reagent C (PBS). Mix thoroughly. (Alternatively, prepare proportionally as needed.)     4) The absorbance (A500) of this solution in a 1cm cuvette should be 0.9–1.3 at 500nm.     5) The mixed solution is stable for two weeks at 4°C. If precipitation occurs in the HABA solution, filter or centrifuge before use.  **Choose either the spectrophotometric method or the microplate assay to measure absorbance at 500nm.**  ### **I. Spectrophotometric Method**  1. Add 900µL of HABA-Avidin solution to a 1cm cuvette and measure the absorbance at 500nm (A1\u003csub\u003e500\u003c\/sub\u003e).  2. Add 100µL of biotin-labeled protein sample to the cuvette containing HABA-Avidin and mix well.  3. Measure the absorbance at 500nm. If the value is ≥0.3 and remains stable for at least 15 seconds, record it as A2\u003csub\u003e500\u003c\/sub\u003e. If the value is \u0026lt;0.3, dilute the biotin-labeled protein sample and repeat the measurement, accounting for the dilution factor in subsequent calculations.  ### **Coupling Efficiency Calculation**  According to the Beer-Lambert Law: A\u003csub\u003eλ\u003c\/sub\u003e = ε\u003csub\u003eλ\u003c\/sub\u003ebC, where:  - **A**: Absorbance at wavelength λ (500nm for HABA).  - **ε**: Molar absorptivity at λ (34000 M\u003csup\u003e-1\u003c\/sup\u003ecm\u003csup\u003e-1\u003c\/sup\u003e for HABA-avidin at pH 7.0).  - **b**: Path length (cm). For a 1cm cuvette, b = 1cm.  - **C**: Molar concentration (mol\/L = mmol\/mL).  **Steps:**  1. Calculate protein molar concentration:     Protein (mmol\/mL) = Protein concentration (mg\/mL) \/ Protein molecular weight (mg\/mmol).  2. Calculate ΔA\u003csub\u003e500\u003c\/sub\u003e:     ΔA\u003csub\u003e500\u003c\/sub\u003e = 0.9 × A1\u003csub\u003e500\u003c\/sub\u003e – A2\u003csub\u003e500\u003c\/sub\u003e     *(Note: 0.9 is a correction factor for dilution by the biotin-labeled protein sample.)*  3. Calculate biotin molar concentration:     Biotin (mmol\/mL) = ΔA\u003csub\u003e500\u003c\/sub\u003e \/ 34000 *(for 1cm cuvette)*.  4. Calculate biotin-to-protein ratio:     Biotin:Protein (molar ratio) = [Biotin (mmol\/mL) × 10] \/ [Protein (mmol\/mL) × dilution factor].  **Example Calculation**  For a biotin-labeled antibody (MW = 150,000, concentration = 3.0mg\/mL), A1\u003csub\u003e500\u003c\/sub\u003e = 1.0, A2\u003csub\u003e500\u003c\/sub\u003e = 0.56:  - Protein molar concentration: 3 \/ 150,000 = 2×10\u003csup\u003e-5\u003c\/sup\u003e mmol\/mL.  - ΔA\u003csub\u003e500\u003c\/sub\u003e: 0.9 × 1.0 – 0.56 = 0.34.  - Biotin molar concentration: 0.34 \/ 34000 = 1×10\u003csup\u003e-5\u003c\/sup\u003e mmol\/mL.  - Biotin:Protein ratio = (1×10\u003csup\u003e-5\u003c\/sup\u003e × 10) \/ (2×10\u003csup\u003e-5\u003c\/sup\u003e) = 5:1.  ### **II. Microplate Assay**  1. Add 180µL of HABA-Avidin solution to a microplate well and measure absorbance at 500nm (A1\u003csub\u003e500\u003c\/sub\u003e).  2. Add 20µL of biotin-labeled protein sample to the well and mix well.  3. Measure absorbance at 500nm. Record the stable value as A2\u003csub\u003e500\u003c\/sub\u003e (after ≥15 seconds).  ### **Coupling Efficiency Calculation**  Beer-Lambert Law applies similarly, with:  - **b**: Path length = 0.59cm (for 200µL in the provided 96-well microplate).  **Steps:**  1. Protein molar concentration: Same as above.  2. ΔA\u003csub\u003e500\u003c\/sub\u003e = A1\u003csub\u003e500\u003c\/sub\u003e – A2\u003csub\u003e500\u003c\/sub\u003e *(no correction factor needed for microplate)*.  3. Biotin molar concentration:     Biotin (mmol\/mL) = ΔA\u003csub\u003e500\u003c\/sub\u003e \/ (34000 × 0.59).  4. Biotin-to-protein ratio: Same formula as above.  **Example Calculation**  For the same antibody (A1\u003csub\u003e500\u003c\/sub\u003e = 0.9, A2\u003csub\u003e500\u003c\/sub\u003e = 0.56):  - ΔA\u003csub\u003e500\u003c\/sub\u003e: 0.9 – 0.56 = 0.34.  - Biotin molar concentration: 0.34 \/ (34000 × 0.59) = 1.7×10\u003csup\u003e-5\u003c\/sup\u003e mmol\/mL.  - Biotin:Protein ratio = (1.7×10\u003csup\u003e-5\u003c\/sup\u003e × 10) \/ (2×10\u003csup\u003e-5\u003c\/sup\u003e) = 8.5:1.  ---\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"Kit","offer_id":41738024583243,"sku":"UA080468-Kit","price":120.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_1cfd5638-d4c5-4465-a47f-ea00da3b8d8f.png?v=1789113620"},{"product_id":"biotin-cleavable-rapid-coupling-kit-ua080467","title":"Biotin (cleavable) Rapid Coupling Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eBiotin (Cleavable) Rapid Conjugation Kit\u003c\/td\u003e\n\u003c\/tr\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eThe Biotin (Biotin, Cleavable) Rapid Conjugation Kit provides a simple and fast conjugation protocol for coupling biotin derivatives (Biotin-S-S) to antibodies, proteins, peptides, or other ligands containing free amino groups. The reagents effectively react with primary amines (-NH2) in buffers at pH 7-9 to form stable amide bonds. Proteins, including antibodies, typically possess several primary amines on the side chains of lysine (K) residues and at the N-terminus of each polypeptide, which can serve as targets for reagent labeling. The conjugation agent contains a disulfide bond in its linear chain, which can be cleaved by reducing agents such as DTT, BME, or TCEP, thereby removing the biotin label and providing additional functionality to the reagent. For example, it can be used for purifying antibodies or proteins, cleaving interactions with streptavidin probes, or serving as a cleavable ADC linker for synthesizing antibody-drug conjugates (ADCs).\u003cbr\u003e\u003cbr\u003eApplications\u003cbr\u003e(1) Rapid labeling of proteins (antibodies), etc.\u003cbr\u003e \u003cbr\u003eAdvantages\u003cbr\u003e(1) High sensitivity and labeling efficiency;\u003cbr\u003e(2) Includes a desalting column and usage protocol;\u003cbr\u003e(3) Provides a method for detecting labeling efficiency;\u003cbr\u003e(4) This kit offers a complete set of reagents for rapid labeling, and the reagents are easy to store.\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e2mg can be used to label 0.5-2mg antibodies (mAb, pAb) or other proteins:\u003c\/p\u003e\n\u003ctable style=\"width: 698.41px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 155.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 155.009px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eVolume\/Amount\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 160.009px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eStorage Temperature\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 227.009px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eRemarks\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent A\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e200µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eAntibody\/Protein Diluent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent B\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e250µg\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCrosslinker (Lyophilized Powder)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e100µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCo-solvent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent D\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e60mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003ePBS Concentrate (20X)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent E\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e14.3mg\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReducing Agent (TCEP.HCl, Lyophilized Powder)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent F\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReducing Agent Buffer\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eDesalting Column\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1 piece\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e10mg can be used to label 0.5-10mg antibodies (mAb, pAb) or other proteins:\u003c\/p\u003e\n\u003ctable style=\"width: 702.41px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 158.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 146.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eVolume\/Amount\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 168.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eStorage Temperature\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 229.007px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eRemarks\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent A\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eAntibody\/Protein Diluent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent B\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1.1mg\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCrosslinker (Lyophilized Powder)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e300µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCo-solvent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent D\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e60mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003ePBS Concentrate (20X)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent E\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e28.6mg\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReducing Agent (TCEP.HCl, Lyophilized Powder)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent F\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReducing Agent Buffer\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eDesalting Column\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1 piece\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cimg class=\"image_resized img img-fluid o_we_custom_image\" style=\"aspect-ratio:1003\/151;width:59.35%;\" src=\"https:\/\/univ-pmc.oss-cn-shanghai.aliyuncs.com\/product.template\/20090828\/8c4a6d82-60a1-4d8a-b854-b158ade2150b.png?access_token=40d38636-2fc1-41de-930a-8033b051c30b\" alt=\"\" width=\"1003\" height=\"151\"\u003e\u003cbr\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eUsage Instructions for 2mg Kit\u003c\/strong\u003e\u003cbr\u003e1. Add Reagent A to the antibody or protein to be labeled (volume ratio of protein to be labeled to Reagent A is 9:1), pipette up and down repeatedly to mix thoroughly, avoiding bubble formation;\u003cbr\u003e2. Add Reagent C (50 µL) to the Reagent B tube, dissolve and mix well to prepare a 5 mg\/mL coupling agent solution (prepare fresh and use immediately);\u003cbr\u003e3. Continue to add the freshly prepared coupling agent to the mixture from Step 1 (add 10 µL of coupling agent solution per 0.5 mg of protein\/antibody; if the amount of protein\/antibody to be labeled increases, increase the coupling agent dosage proportionally), pipette up and down to mix thoroughly, avoiding bubble formation;\u003cbr\u003e4. React at room temperature for 2 hours, or mix slowly on a mixer;\u003cbr\u003e5. Dilute Reagent D concentrate 20-fold with distilled or purified water to prepare column buffer and dialysis buffer.\u003cbr\u003e6. Desalting purification: ① Equilibrate the desalting column with the diluted Reagent D solution, using an equilibration volume of 30-40 mL. ② Adjust the volume of the conjugated protein\/antibody to 2.5 mL with column buffer, load it onto the desalting column, allow it to drain completely, and do not collect the flow-through. ③ Add another 3.5 mL of column buffer to the desalting column and collect 3.5 mL of flow-through.\u003cbr\u003e7. Dialysis (optional): The reaction mixture can be dialyzed against PBS dialysis buffer (10 mM, pH 7.2-7.4) prepared by diluting Reagent D, or other dialysis buffers, at 2-8°C for 24-36 hours, changing the buffer 3-4 times. During dialysis, the buffer should be gently stirred.\u003cbr\u003e8. Cleave the disulfide bond in the spacer arm: ① Reducing agent preparation: Add Reagent F (1 mL) to the Reagent E tube, dissolve and mix well to prepare a 50 mM TCEP·HCl reducing agent solution, pH 5.5. ② Incubate the sample at room temperature for 30 minutes. ③ Alternatively, the sample can be incubated in 50 mM DTT or 2-mercaptoethanol at room temperature for 2 hours and 30 minutes.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eUsage Instructions for 10mg Kit\u003c\/strong\u003e\u003cbr\u003e1. Add Reagent A to the antibody or protein to be labeled (volume ratio of protein to be labeled to Reagent A is 9:1), pipette up and down repeatedly to mix thoroughly, avoiding bubble formation;\u003cbr\u003e2. Add Reagent C (220 µL) to the Reagent B tube, dissolve and mix well to prepare a 5 mg\/mL coupling agent solution (prepare fresh and use immediately);\u003cbr\u003e3. Continue to add the freshly prepared coupling agent to the mixture from Step 1 (add 10 µL of coupling agent solution per 0.5 mg of protein\/antibody; if the amount of protein\/antibody to be labeled increases, increase the coupling agent dosage proportionally), pipette up and down to mix thoroughly, avoiding bubble formation;\u003cbr\u003e4. React at room temperature for 2 hours, or mix slowly on a mixer;\u003cbr\u003e5. Dilute Reagent D concentrate 20-fold with distilled or purified water to prepare column buffer and dialysis buffer.\u003cbr\u003e6. Desalting purification: ① Equilibrate the desalting column with the diluted Reagent D solution, using an equilibration volume of 30-40 mL. ② Adjust the volume of the conjugated protein\/antibody to 2.5 mL with column buffer, load it onto the desalting column, allow it to drain completely, and do not collect the flow-through. ③ Add another 3.5 mL of column buffer to the desalting column and collect 3.5 mL of flow-through.\u003cbr\u003e7. Dialysis (optional): The reaction mixture can be dialyzed against PBS dialysis buffer (10 mM, pH 7.2-7.4) prepared by diluting Reagent D, or other dialysis buffers, at 2-8°C for 24-36 hours, changing the buffer 3-4 times. During dialysis, the buffer should be gently stirred.\u003cbr\u003e8. Cleave the disulfide bond in the spacer arm: ① Reducing agent preparation: Add Reagent F (2 mL) to the Reagent E tube, dissolve and mix well to prepare a 50 mM TCEP·HCl reducing agent solution, pH 5.5. ② Incubate the sample at room temperature for 30 minutes. ③ Alternatively, the sample can be incubated in 50 mM DTT or 2-mercaptoethanol at room temperature for 2 hours and 30 minutes.① Reducing agent preparation: Add Reagent F (2 mL) to the Reagent E tube, dissolve and mix well to prepare a 50 mM TCEP·HCl reducing agent solution, pH 5.5. ② Incubate the sample at room temperature for 30 minutes. ③ Alternatively, the sample can be incubated in 50 mM DTT or 2-mercaptoethanol at room temperature for 2 hours and 30 minutes.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eInstructions for Gravity Desalting Column\u003c\/strong\u003e\u003cbr\u003e1. Preparation before using the gravity desalting column\u003cbr\u003e1). Remove the top cap of the desalting column and pour out the column storage solution;\u003cbr\u003e2). Cut or snip off the sealed end at the bottom of the column.\u003cbr\u003e2. Desalting purification\u003cbr\u003e1). Equilibrate the desalting column with a 20-fold dilution of Reagent D or other buffers, using an equilibration volume of 35-40 mL, and discard the eluate;\u003cbr\u003e2). Adjust the volume of the conjugated protein\/antibody to 2.5 mL with column buffer, load it onto the desalting column, allow it to drain completely, and do not collect the flow-through;\u003cbr\u003e3). Add another 3.5 mL of column buffer to the desalting column and collect 3-3.5 mL of flow-through (EP tubes can be used, collecting 1 mL per tube);\u003cbr\u003e3. Operating conditions and regeneration\/storage of the desalting column\u003cbr\u003e1). Operating conditions for the desalting column: pH 2-13, 4°C -30°C; stable in common buffers (PBS, Tris-HCl, sodium carbonate-sodium bicarbonate, etc.);\u003cbr\u003e2). Recommended for single use (can also be regenerated and reused);\u003cbr\u003e3). Regeneration and storage of the desalting column: After use, wash thoroughly and store the column in 20% ethanol, 0.05% NaN3, or 0.05 mol\/L NaOH.\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e1. Buffer and concentration requirements for antibodies\/proteins to be conjugated: Antibodies\/proteins should be stored in 10 mM phosphate buffer (1x PBS, pH 7.2-7.4). The recommended concentration range is 2-4 mg\/mL. The buffer must not contain amine components to avoid competitive reactions with the coupling reagent, which may affect conjugation efficiency. This includes primary amines (e.g., Tris, glycine, ammonium salts, EDTA) and protein stabilizers (e.g., BSA, gelatin). If the sample contains substances that may interfere with labeling, it is recommended to exchange the buffer with PBS and concentrate it to the recommended concentration.2. For most IgGs, a 1 mg\/mL solution in a 1 cm pathlength cuvette has an absorbance of approximately 1.3-1.4 at 280 nm. The coupling reagent does not exhibit significant absorption at 280 nm. Therefore, the concentration (mg\/mL) of the biotin-labeled antibody can be calculated by dividing the absorbance value of the dialyzed sample at 280 nm by 1.4.3. It is recommended to store the conjugated product under refrigeration. If the final concentration of the purified antibody\/protein conjugate is less than 1 mg\/mL, adding bovine serum albumin (BSA) or other protein stabilizers to a concentration of 1-10 mg\/mL can stabilize it for several months at 2-8°C. For long-term storage, the labeled solution can be aliquoted as needed and stored at -20°C in the dark, avoiding repeated freeze-thaw cycles.4. The coupling reagent solution prepared with the solubilizing agent included in this kit should be sealed and stored at -20°C if not fully used. It can be used normally within one week. If used within one month, the amount should be increased appropriately. Do not use it beyond one month.5. For small peptide molecules with a molecular weight of less than 5 kDa, the amount of coupling reagent solution can be appropriately increased to improve conjugation efficiency.6. Instructions for using the desalting column (\u0026gt;5000 Mr) included in this kit: column volume 8.3 mL, elution volume 3.5 mL. For gravity desalting: sample loading volume 1.0-2.5 mL, desalting efficiency \u0026gt;98%. For centrifugal desalting: sample loading volume 1.75-2.5 mL, desalting efficiency \u0026gt;90%.7. For Reagent D, salt crystallization may occur when stored at 2°C-8°C. This is a normal phenomenon, and the crystals will disappear after returning to room temperature and mixing.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"2mg","offer_id":41738024616011,"sku":"UA080467-2mg","price":370.0,"currency_code":"USD","in_stock":true},{"title":"10mg","offer_id":41738024648779,"sku":"UA080467-10mg","price":900.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_946040d2-12e4-4c2f-85e8-31dba75cbf8d.png?v=1789113617"},{"product_id":"biotin-rapid-coupling-kit-ua080466","title":"Biotin Rapid Coupling Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eBiotin Rapid Conjugation Kit\u003c\/td\u003e\n\u003c\/tr\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eThe Biotin Rapid Conjugation Kit provides a simple and fast conjugation protocol for coupling biotin to antibodies, proteins, peptides, or other ligands containing free amino groups. The reagents effectively react with primary amines (-NH\u003csub\u003e2\u003c\/sub\u003e) in buffers at pH 7-9, forming stable amide bonds. Proteins, including antibodies, typically possess several primary amines on the side chains of lysine (K) residues and at the N-terminus of each polypeptide, which can serve as targets for reagent labeling.\u003cbr\u003e\u003cbr\u003eApplications\u003cbr\u003e(1) Rapid labeling of proteins (antibodies), etc.\u003cbr\u003e \u003cbr\u003eAdvantages\u003cbr\u003e(1) High sensitivity and labeling efficiency;\u003cbr\u003e(2) Includes a desalting column and usage protocol;\u003cbr\u003e(3) Provides a method for detecting labeling efficiency;\u003cbr\u003e(4) This kit offers a complete set of reagents for rapid labeling, and the reagents are easy to store.\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e2mg can be used to label 0.5-2mg antibodies (monoclonal, polyclonal) or other proteins:\u003cbr\u003e\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 694.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 138.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2mg Components\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 131.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eVolume\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 200.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eStorage Temperature\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 224.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eRemarks\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent A\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e200µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eAntibody\/Protein Diluent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent B\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e200µg\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCrosslinker (Lyophilized Powder)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e100µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCo-solvent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent D\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e60mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003ePBS Concentrate (20X)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eDesalting Column\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1 column\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e10mg can be used to label 0.5-10mg antibodies (monoclonal, polyclonal) or other proteins:\u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 678.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 143.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e10mg Components\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 124.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eVolume\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 197.007px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eStorage Temperature\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 213.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eRemarks\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent A\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eAntibody\/Protein Diluent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent B\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1mg\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCrosslinker (Lyophilized Powder)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e300µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCo-solvent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent D\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e60mL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003ePBS Concentrate (20X)\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eDesalting Column\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e1 column\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e2°C-8°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cimg alt=\"\" class=\"img img-fluid o_we_custom_image image_resized\" height=\"151\" src=\"https:\/\/univ-pmc.oss-cn-shanghai.aliyuncs.com\/product.template\/20090829\/e1167336-f3a6-4545-b9f4-23b2566193e2.png?access_token=edc449ee-0794-4312-beca-d870b1522e19\" style=\"aspect-ratio:1003\/151;width:54.84%;\" width=\"1003\"\u003e\u003cbr\u003e\u003cspan\u003e\u003cstrong\u003e2mg Usage Instructions\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e1. This reagent is sensitive to moisture. Before opening, ensure that each vial of the components is fully equilibrated to room temperature to prevent condensation inside the container.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e2. Add Reagent A to the antibody or protein to be labeled (the volume ratio of the protein to Reagent A is 9:1), and pipette up and down repeatedly to mix thoroughly, avoiding the formation of bubbles.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e3. Add Reagent C (40 µL) to the Reagent B tube, dissolve and mix well to prepare a 5 mg\/mL coupling agent solution (prepare fresh and use immediately).\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e4. To the mixture from Step 2, add the freshly prepared coupling agent (add 8 µL of Reagent B solution for every 0.5 mg of protein\/antibody; if the amount of protein\/antibody being labeled increases, proportionally increase the amount of coupling agent added). Mix by pipetting up and down, avoiding the formation of bubbles.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e5. React at room temperature for 2 hours, or gently mix on a shaker;\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e6. Dilute the concentrated Reagent D 20-fold with distilled or purified water to prepare the column buffer and dialysis solution;\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e7. Desalt using a desalting column (see instructions below); load 2.5 mL of sample, and collect the filtered protein solution in collection tubes—this is the conjugation product.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e8. Dialysis (optional): Use PBS dialysis buffer prepared by diluting Reagent D (10 mM, pH 7.2–7.4) or another suitable dialysis buffer, and dialyze the reaction mixture at 2–8°C for 24–36 hours, changing the buffer 3–4 times. Gently stir the buffer during dialysis.\u003c\/span\u003e\u003cbr\u003e\u003cbr\u003e\u003cspan\u003e\u003cstrong\u003eUsage instructions for 10 mg\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e1. This reagent is sensitive to moisture. Before opening, ensure that each vial of the components is fully equilibrated to room temperature to prevent condensation inside the container.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e2. Add Reagent A to the antibody or protein to be labeled (the volume ratio of the protein to be labeled to Reagent A is 9:1), and pipette up and down repeatedly to mix thoroughly, avoiding the formation of bubbles.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e3. Add Reagent C (200 µL) to the Reagent B tube, dissolve and mix well to prepare a 5 mg\/mL coupling agent solution (prepare fresh before use).\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e4. Continue by adding the freshly prepared coupling agent to the mixture from Step 2 (add 8 µL of Coupling Agent B solution for every 0.5 mg of protein\/antibody; if the amount of protein\/antibody being labeled increases, proportionally increase the amount of coupling agent added). Pipette up and down to mix thoroughly, avoiding the formation of bubbles.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e5. Allow the reaction to proceed at room temperature for 2 hours, or gently mix on a shaker.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e6. Dilute reagent D concentrate 20-fold with distilled or purified water to prepare the column buffer and dialysis solution.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e7. Perform desalting using a desalting column (see instructions below); load 2.5 mL of sample, and collect the filtered protein solution in collection tubes—this is the conjugation product.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e8. Dialysis (optional): Use PBS dialysis buffer prepared by diluting reagent D (10 mM, pH 7.2–7.4) or another suitable dialysis buffer, and dialyze the reaction mixture at 2–8°C for 24–36 hours, changing the buffer 3–4 times. During dialysis, stir the buffer gently.\u003c\/span\u003e\u003cbr\u003e\u003cbr\u003e\u003cspan\u003e\u003cstrong\u003eInstructions for Use of the Gravity Desalting Column\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e1. Preparations Before Using the Gravity Desalting Column\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e1). Remove the top cap of the desalting column and pour out the storage solution;\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e2). Cut or trim off the sealed end at the bottom of the column.\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e2. Desalting purification\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e1). Equilibrate the desalting column with a 20-fold dilution of Reagent D or another buffer, using an equilibration volume of 35–40 mL, and discard the eluate;\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e2). Adjust the volume of the post-coupling protein\/antibody to 2.5 mL with column buffer, add it to the desalting column, allow it to drain completely, and do not collect the filtrate;\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e3). Add 3.5 mL of column buffer to the desalting column again, while collecting 3–3.5 mL of the filtrate (using EP tubes, collecting 1 mL per tube);\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e3. Operating conditions and regeneration\/storage of the desalting column\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e1). Working conditions of the desalting column: pH 2–13, 4°C–30°C; stable in commonly used buffer solutions (PBS, Tris-HCl, sodium carbonate–sodium bicarbonate, etc.).\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e2). Single-use is recommended (may also be reused after regeneration).\u003c\/span\u003e\u003cbr\u003e\u003cspan\u003e3). Regeneration and storage of the desalting column: After passing the sample through the column, wash thoroughly, then store the column in 20% ethanol, 0.05% NaN3, or 0.05 mol\/L NaOH.\u003c\/span\u003e\u003cbr\u003e \u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e1. Requirements for the antibody\/protein buffer to be conjugated and concentration: The antibody\/protein should be stored in 10 mM phosphate buffer (1x PBS, pH 7.2-7.4). The recommended concentration range is 2-4 mg\/mL. The buffer must not contain amino groups to avoid competitive reactions with the coupling reagent, which may affect conjugation efficiency. Specifically, primary amines (e.g., Tris, glycine, ammonium salts, EDTA) and protein stabilizers (e.g., BSA, gelatin) should be avoided. If the sample contains substances that may interfere with labeling, it is recommended to exchange the buffer with PBS and concentrate to the recommended concentration.  2. For most IgGs, a 1 mg\/mL solution in a 1 cm pathlength cuvette has an absorbance of approximately 1.3-1.4 at 280 nm. The coupling reagent does not exhibit significant absorption at 280 nm. Therefore, the concentration (mg\/mL) of the biotin-labeled antibody can be calculated by dividing the absorbance of the dialyzed sample at 280 nm by 1.4.  3. It is recommended to store the conjugated product under refrigeration. If the final concentration of the purified antibody\/protein conjugate is less than 1 mg\/mL, adding bovine serum albumin (BSA) or another protein stabilizer to a concentration of 1-10 mg\/mL can stabilize it for several months at 2-8°C. For long-term storage, the labeled solution can be aliquoted as needed and frozen at -20°C in the dark, avoiding repeated freeze-thaw cycles.  4. The coupling reagent solution prepared with the solubilizing agent included in this kit should be sealed and stored at -20°C if unused. It can be used normally within one week. If stored for more than one week but within one month, the amount used should be increased appropriately. Do not use if stored for more than one month.  5. For small peptide molecules with a molecular weight of less than 5 kDa, the amount of coupling reagent solution added can be appropriately increased to improve conjugation efficiency.  6. Instructions for use of the desalting column (\u0026gt;5000 Mr) included in this kit: Column volume 8.3 mL, elution volume 3.5 mL. Gravity desalting: Sample loading volume 1.0-2.5 mL, desalting efficiency \u0026gt;98%. Centrifugal desalting: Sample loading volume 1.75-2.5 mL, desalting efficiency \u0026gt;90%.  7. Reagent D may form salt crystals when stored at 2°C -8°C. This is normal; the crystals will disappear after returning to room temperature and mixing.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"2mg","offer_id":41738024681547,"sku":"UA080466-2mg","price":320.0,"currency_code":"USD","in_stock":true},{"title":"10mg","offer_id":41738024714315,"sku":"UA080466-10mg","price":800.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_53f19b3a-c895-4203-b28d-78d0f1f3c56b.png?v=1789106660"},{"product_id":"horseradish-peroxidase-hrp-rapid-coupling-kit-ua080470","title":"Horseradish peroxidase (HRP) Rapid Coupling Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eRapid HRP Conjugation Kit\u003c\/td\u003e\n\u003c\/tr\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003eThis kit provides a simple and rapid conjugation solution specifically designed for coupling horseradish peroxidase (HRP) to antibodies, proteins, peptides, or other ligands containing free amino groups. Compared to traditional methods, our conjugation reaction offers higher efficiency and yield, with more stable conjugates that are less prone to detachment. The reagents effectively react with primary amines (-NH2) in buffers at pH 7-9, forming stable amide bonds. Proteins, including antibodies, typically possess several primary amines on the side chains of lysine (K) residues and at the N-terminus of each polypeptide, which serve as targets for reagent labeling.\u003cbr\u003e\u003cbr\u003eFeatures and Benefits:\u003cbr\u003eThe kit provides a complete set of reagents required for rapid labeling, and the reagents are easy to store.\u003cbr\u003e100% recovery rate, with a conjugation process completed in 2 hours and hands-on time of approximately 5 minutes.\u003cbr\u003eThe conjugated products require no purification and can be stored for extended periods.\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e100μg can be used to label 100µg of antibody (monoclonal, polyclonal) or other proteins:\u003c\/p\u003e\n\u003ctable style=\"width: 639.41px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 127.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 114.007px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eVolume\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 233.002px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          Storage Temperature\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 164.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eRemarks\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent A\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e20µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eAntibody\/Protein Diluent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent B\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e10µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C Store protected from light\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCrosslinker\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e15µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C Store protected from light\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eActivated Enzyme\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e500μg can be used to label 500µg of antibody (monoclonal, polyclonal) or other proteins:\u003c\/p\u003e\n\u003ctable style=\"height: 218.602px; width: 639.41px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 54.7107px;\"\u003e\n\u003ctd style=\"width: 123.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 115.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eVolume\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 236.012px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          Storage Temperature\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 164.005px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eRemarks\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 54.7107px;\"\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent A\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e50µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eAntibody\/Protein Diluent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 54.7107px;\"\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent B\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e30µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C Store protected from light\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCrosslinker\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 53.7292px;\"\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e60µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C Store protected from light\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eActivated Enzyme\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e1mg can be used to label 1mg of antibody (monoclonal, polyclonal) or other proteins:\u003c\/p\u003e\n\u003ctable style=\"width: 639.41px;\" class=\"table table-bordered\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 122.669px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eComponent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 118.007px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eVolume\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 231.009px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          Storage Temperature\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 167.002px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eRemarks\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent A\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e100µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eAntibody\/Protein Diluent\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent B\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e60µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C Store protected from light\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eCrosslinker\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eReagent C\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e110µL\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003e          -20°C Store protected from light\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 298.218px;\"\u003e\u003cp\u003e\u003cspan style=\"color:rgb(0,0,0);font-size:9.0000pt\"\u003e\u003cfont\u003eActivated Enzyme\u003c\/font\u003e\u003c\/span\u003e\u003cspan style=\"font-size:12.0000pt\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cstrong\u003e100μg Usage Method:\u003c\/strong\u003e\u003cbr\u003e1. Add Reagent A to the antibody or protein to be labeled (concentration range: 3-10mg\/mL) (volume ratio of protein to Reagent A is 9:1), mix thoroughly by pipetting repeatedly, avoiding bubbles;\u003cbr\u003e2. Continue by adding Reagent B (5µL) to the antibody or protein to be labeled, mix thoroughly by pipetting repeatedly; avoid bubbles;\u003cbr\u003e3. Incubate at room temperature in the dark with gentle mixing for 1 hour;\u003cbr\u003e4. Continue by adding Reagent C (10µL), mix thoroughly by pipetting repeatedly; avoid bubbles.\u003cbr\u003e   Note: Depending on the protein or antibody, the amount of Reagent C may be adjusted as needed.\u003cbr\u003e5. Incubate at room temperature in the dark with gentle mixing for 1 hour.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e500μg Usage Method\u003c\/strong\u003e\u003cbr\u003e1. Add Reagent A to the antibody or protein to be labeled (concentration range: 3-10mg\/mL) (volume ratio of protein to Reagent A is 9:1), mix thoroughly by pipetting repeatedly, avoiding bubbles;\u003cbr\u003e2. Continue by adding Reagent B (25µL) to the antibody or protein to be labeled, mix thoroughly by pipetting repeatedly; avoid bubbles;\u003cbr\u003e3. Incubate at room temperature in the dark with gentle mixing for 1 hour;\u003cbr\u003e4. Continue by adding Reagent C (50µL), mix thoroughly by pipetting repeatedly; avoid bubbles.\u003cbr\u003e   Note: Depending on the protein or antibody, the amount of Reagent C may be adjusted as needed.\u003cbr\u003e5. Incubate at room temperature in the dark with gentle mixing for 1 hour.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e1mg Usage Method\u003c\/strong\u003e\u003cbr\u003e1. Add Reagent A to the antibody or protein to be labeled (concentration range: 3-10mg\/mL) (volume ratio of protein to Reagent A is 9:1), mix thoroughly by pipetting repeatedly, avoiding bubbles;\u003cbr\u003e2. Continue by adding Reagent B (50µL) to the antibody or protein to be labeled, mix thoroughly by pipetting repeatedly; avoid bubbles;\u003cbr\u003e3. Incubate at room temperature in the dark with gentle mixing for 1 hour;\u003cbr\u003e4. Continue by adding Reagent C (100µL), mix thoroughly by pipetting repeatedly; avoid bubbles.\u003cbr\u003e   Note: Depending on the protein or antibody, the amount of Reagent C may be adjusted as needed.\u003cbr\u003e5. Incubate at room temperature in the dark with gentle mixing for 1 hour.\u003cbr\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e1. This reagent is sensitive to moisture. Before opening the vial, ensure all component vials are fully equilibrated to room temperature to avoid moisture condensation inside the container.  2. Requirements for the antibody\/protein to be conjugated: The antibody\/protein should be stored in a 10 mM phosphate buffer (1x PBS, pH 7.2-7.4). The recommended concentration range is 3-10 mg\/mL. The buffer must not contain any amine groups to avoid competitive reactions with the coupling reagent, which may affect conjugation efficiency. Specifically, avoid primary amines (e.g., Tris, glycine, ammonium salts, EDTA), protein stabilizers (e.g., BSA \u0026lt;0.1%, gelatin, glycerol \u0026lt;20%), and preservatives such as sodium azide. If the sample contains substances that may interfere with labeling, it is recommended to exchange the buffer with PBS and concentrate to the recommended concentration.  3. Repeated freeze-thaw cycles significantly impact this reagent. Any unused thawed reagent should be discarded and not reused.  4. Storage of conjugated products: If the final concentration of the antibody\/protein conjugate is less than 0.5 mg\/mL, consider concentrating or adding stabilizers. Store at 2-8°C with a shelf life of 3 months. For long-term storage, the conjugated product can be lyophilized or stored at -20°C after adding 50% glycerol.\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"100μg","offer_id":41738025697355,"sku":"UA080470-100μg","price":370.0,"currency_code":"USD","in_stock":true},{"title":"500μg","offer_id":41738025730123,"sku":"UA080470-500μg","price":870.0,"currency_code":"USD","in_stock":true},{"title":"1mg","offer_id":41738025762891,"sku":"UA080470-1mg","price":1085.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_269a510a-454e-4237-acb9-26631ec1ff54.png?v=1789113615"},{"product_id":"antibody-glycan-site-specific-conjugation-kit-ua079023","title":"Antibody Glycan Site-specific Conjugation Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"color: rgb(102, 102, 102)\"\u003eThe antibody site-specific conjugation kit developed based on antibody glycan editing technology utilizes the N-297 conserved glycan in the CH2 region of the IgG heavy chain of the antibody to mediate site-specific conjugation. The site-specific modification is achieved using glycosidase and glycosyltransferase.\u003cbr\u003e\u003cbr\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"color: rgb(102, 102, 102)\"\u003eThis kit enables rapid site-specific conjugation of monoclonal antibodies, overcoming the drawbacks of conventional conjugation methods such as random conjugation sites, heterogeneous products, or the need for antibody engineering. This kit provides a complete set of reaction reagents for antibody azidation modification and can be completed within 2 days. \u003cbr\u003e \u003cbr\u003eAs shown in the figure below, first, the sugarase EndoS is used to expose the acetylgalactosamine (blue square) on the conserved N-glycan of the constant region of the monoclonal antibody. Then, the mutant of bovine galactosyltransferase (b.GalTY298L) is used to link the acetylated galactosamine with azide functional groups (yellow square - N3) to the acetylgalactosamine. Subsequently, the copper-free catalyzed acetylenecyanide cycloaddition reaction (such as SPAAC) can be used to link biotin, fluorescein or toxin molecules, etc. to the monoclonal antibody.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003cimg src=\"https:\/\/univ-pmc.oss-cn-shanghai.aliyuncs.com\/product.template\/20152858\/dfca9b35-3b54-44a2-bf16-58b3627864d6.png?access_token=751c973c-7963-4383-b87b-4a0cd4726123\" class=\"img-fluid img o_we_custom_image\"\u003e\u003cimg src=\"https:\/\/univ-pmc.oss-cn-shanghai.aliyuncs.com\/product.template\/20152858\/087fe850-4ebe-492c-ad2f-97d588c77724.png?access_token=c3f6d7bb-6f29-4a15-92e7-1992ab5bd937\" class=\"img-fluid img o_we_custom_image\"\u003e\u003c\/p\u003e\n\u003cp\u003eProduct features:\u003cbr\u003eNo need to modify the antibody, without damaging the antibody structure.\u003cbr\u003eThe product is uniform, with high and stable coupling efficiency.\u003cbr\u003eAntibodies with glycosylation modification can all be applied, and they are compatible with various linkers and drugs.\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cfont class=\"text-black\"\u003eS Size：\u003c\/font\u003e \u003cspan style=\"color:rgba(0,0,0,0.9);\"\u003e\u003cfont class=\"text-black\"\u003eLabeling 5 mg of antibody\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cfont class=\"text-black\"\u003eM Size：\u003c\/font\u003e\u003cspan style=\"color:rgba(0,0,0,0.9);\"\u003e\u003cfont class=\"text-black\"\u003eLabeling 10 mg of antibody\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cfigure class=\"table\"\u003e\u003ctable style=\"border-collapse:collapse;width:546.5pt;\" border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"729\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height:20.25pt;\"\u003e\u003ctd style=\"border:solid black 1.0pt;height:20.25pt;padding:0cm 5.4pt 0cm 5.4pt;width:546.5pt;\" colspan=\"4\" width=\"729\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e\u003cstrong\u003eSize: 1 mg(For labelling 1 mg antibody)\u003c\/strong\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\u003c\/tr\u003e\n\u003ctr style=\"height:19.5pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:19.5pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e\u003cstrong\u003eMaterial\u003c\/strong\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:19.5pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e\u003cstrong\u003eAmount\u003c\/strong\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:19.5pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e\u003cstrong\u003eStorage\u003c\/strong\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:19.5pt;padding:0cm 5.4pt 0cm 5.4pt;width:212.15pt;\" width=\"283\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e\u003cstrong\u003eStability\u003c\/strong\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:20.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:20.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eReagentl\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:20.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e5.5\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003eμ\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:20.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e-20\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003e℃\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e,Avoid Freeze Thaw\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:20.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:212.15pt;\" rowspan=\"5\" width=\"283\"\u003e\u003cp style=\"text-align:left;text-indent:20.0pt;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eWhen stored as directed,the componentis stable for 6months.\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:21.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eReagent Il\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e110\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003eμ\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e-20\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003e℃\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e,Avoid Freeze Thaw\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:21.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eReagentIIll\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e110\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003eμ\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e-20\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003e℃\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e,Avoid Freeze Thaw\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:22.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:22.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eReagent IV\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:22.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e11\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003eμ\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:22.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e-20\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003e℃\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:21.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eReagent V\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e5.5\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003eμ\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e-20\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003e℃\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:21.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eSolution A\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e20 mL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eRT\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:212.15pt;\" width=\"283\" valign=\"top\"\u003e \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:21.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eSolution B\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e10 mL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eRT\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:212.15pt;\" rowspan=\"4\" width=\"283\" valign=\"top\"\u003e \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:21.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eSolution C\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e1.5mL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eRT\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:21.0pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eSolution D\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e200\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003eμ\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eRT\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:21.5pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:21.5pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eProteinA Beads\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.5pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e500\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003eμ\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eL\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:21.5pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003e4\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;'\u003e℃\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height:20.25pt;\"\u003e\n\u003ctd style=\"border-top:none;border:solid black 1.0pt;height:20.25pt;padding:0cm 5.4pt 0cm 5.4pt;width:111.9pt;\" width=\"149\"\u003e\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eAC Column\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:20.25pt;padding:0cm 5.4pt 0cm 5.4pt;width:71.5pt;\" width=\"95\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eeach\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:20.25pt;padding:0cm 5.4pt 0cm 5.4pt;width:150.95pt;\" width=\"201\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;font-size:10.0pt;' lang=\"EN-US\"\u003eRT\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid black 1.0pt;border-left:none;border-right:solid black 1.0pt;border-top:none;height:20.25pt;padding:0cm 5.4pt 0cm 5.4pt;width:212.15pt;\" width=\"283\" valign=\"top\"\u003e \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/figure\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003eFor a degree of label (DOL) or antibody-drug ratio (DAR) of 2.0\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003eMaterials Required but Not Provided\u003c\/span\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;'\u003e：\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e● Ultrapure water\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e● DMSO\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e● BCN\/DBCO-toxin\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e● Centrifuge tubes (1.5 mL \/15 mL)\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e● Ultrafiltration spin columns (0.5 mL, 50 kDa MWCO)\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e\u003cstrong\u003eOptimized Protocol for Antibody Azide Labelling\u003c\/strong\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e1.1 Add 5 μL of Reagent I to the antibody solution (10 mg\/mL, 100 μL, in 1×PBS, pH=7.2) and incubate at 37°C for 2 hours. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e1.2 Add 400 μL of 1×PBS (Solution A) to a 50 kDa molecular weight cutoff (MWCO) ultrafiltration tube, then centrifuge at 8,000 × g for 5 minutes at 4°C to wash the ultrafiltration tube. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e1.3 Transfer the sample from step 1.1 into the ultrafiltration tube, then add 300 μL of 2×Tris-HCl buffer (Solution B). Centrifuge under the same conditions as in step 1.2. Repeat this step 5 times, adjusting the final volume to approximately 100 μL. Invert the ultrafiltration tube into a new collection tube and centrifuge at 10,000 × g for 2 minutes at 4°C to collect the sample. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e1.4 In a new 1.5 mL centrifuge tube, sequentially add 285 μL of ultrapure water, 400 μL of Solution B, 100 μL of Reagent II, 100 μL of Reagent III, 10 μL of Reagent IV, and 5 μL of Reagent V. Mix by inversion, then add 100 μL of the sample from above and mix again. Incubate at 30°C in water bath or with rotation for 16 hours. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e(Note: If protein precipitation occurs after the reaction, centrifuge at 10,000 × g for 2 minutes and use the supernatant for subsequent steps.)\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e1.5 Load 500 μL of Protein A beads into an empty column and equilibrate with 3 mL of Solution B. Apply the reaction mixture from step 1.4 onto the beads and allow the sample to flow through by gravity. Collect the flow-through and repeat the loading 3 times to ensure sufficient antibody binding. Wash the beads with 5 mL of Solution A, then elute the antibody with 300 μL of sodium citrate (Solution C). Collect the eluate and repeat the elution 4 times. Combine all elution fractions and slowly add 1 M Tris (Solution D) to adjust the pH to 7.2. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e1.6 Transfer 400 μL of the eluate to a 50 kDa MWCO ultrafiltration tube and centrifuge under the same conditions as in step 1.2. Repeat this step to concentrate all eluates to approximately 100 μL. Add 300 μL of Solution A and centrifuge under the same conditions. Repeat this washing step 5 times. Invert the ultrafiltration tube into a new collection tube and centrifuge at 10,000 × g for 2 minutes at 4°C to collect the sample, yielding the azide-labelled antibody. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e1.7 Determine the concentration of the azide-labelled antibody using NanoDrop or the BCA assay.\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e \u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e\u003cstrong\u003eRecommended Protocol for Toxin Conjugation (Not Included in this Kit)\u003c\/strong\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e2.1 Example Using endo-BCN-PEG4-Val-Cit-PAB-MMAE (CAS: 2762519-08-2): \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003ePrepare a 10 mM stock solution of the toxin molecule in DMSO. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003eAdd the toxin molecule to the azide-labelled antibody at a molar ratio of 1:10 (antibody:toxin). \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003eIncubate at 30°C in water bath or with rotation for 16 hours. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003e2.2 Transfer the reaction product to a 50 kDa molecular weight cutoff ultrafiltration tube. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003eCentrifuge at 8,000 × g for 5 minutes at 4°C. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003eAdd 300 μL of 1× PBS buffer and repeat centrifugation under the same conditions. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003eRepeat the washing step a total of 5 times. \u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\" align=\"left\"\u003e\u003cspan style=\"color:#231916;\"\u003e\u003cspan style='font-family:\"Microsoft YaHei UI\",sans-serif;' lang=\"EN-US\"\u003eInvert the ultrafiltration tube into a new collection tube and centrifuge at 10,000 × g for 2 minutes at 4°C to collect the sample, yielding the MMAE-antibody conjugate.\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"500μg","offer_id":43087187476555,"sku":"UA079023-500μg","price":960.0,"currency_code":"USD","in_stock":true},{"title":"1mg","offer_id":43087187509323,"sku":"UA079023-1mg","price":1620.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_b25af376-c452-480b-be3e-446c19b6a9eb.png?v=1788516140"},{"product_id":"polya-polymerase-tailing-kit-ua070116","title":"Poly(A) Polymerase Tailing Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore at -25 ~ -15℃ for 2 years\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 576.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 233.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e\u003cfont\u003eComponents\u003c\/font\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 161.012px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070116-50 Reactions\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 181.005px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUA070116-250 Reactions\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eE. coli Poly(A) Polymerase\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e(5 U\/µl)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 100 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 X PAP Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e300 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 300 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 mM ATP\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e300 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 300 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase-Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 × 1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e1. Prepare the following reaction system on ice according to the table below:  \u003c\/p\u003e\n\u003ctable class=\"table table-bordered\" style=\"width: 589.41px;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 241.669px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 153.002px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 194.005px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eFinal Concentration\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 X PAP Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 X\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase Inhibitor (40U\/μl)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1.25 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 U\/μl (Optional)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eE.coli Poly(A) Polymerase\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e (5 U\/μl)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e2 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e0.2 U\/μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e10 mM ATP\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e5 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e1 mM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eRNase Free dH2O\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.616px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003eUp to 50 μl\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 397.639px;\"\u003e\u003cp\u003e\u003cspan style=\"font-size:10.5000pt\"\u003e-\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e  Gently pipette to mix thoroughly.  2. Poly(A) tailing reaction: Incubate at 37°C for 30–60 min.  3. Reaction termination: Immediately store at –20°C after completion, or add EDTA to a final concentration of 10 mM, or perform phenol\/chloroform extraction followed by ammonium salt\/ethanol precipitation of RNA.\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e1. Due to the involvement of RNA manipulation, strict adherence to RNA operation protocols is required to avoid RNase contamination. Related reagents and consumables must be treated with DEPC to remove RNase or ensured to be RNase-free.  2. The RNA used for the tailing reaction should be appropriately purified before use and dissolved in RNase-free dH2O. The solution should not contain EDTA or salts.  3. If it is difficult to ensure a strictly RNase-free environment, it is recommended to add an appropriate amount of RNase Inhibitor to the reaction system to enhance the stability of RNA in the solution.  4. During experimental operations, enzyme products should be kept on ice at all times and stored at -20°C immediately after the experiment.\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50Rxns","offer_id":42302149558347,"sku":"UA070116-50Rxns","price":420.0,"currency_code":"USD","in_stock":true},{"title":"50Rxns*5","offer_id":42302149591115,"sku":"UA070116-50Rxns*5","price":1670.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/8115750237f54e8480d50a195a4b4bec.png?v=1788005068"},{"product_id":"baculovirus-immunostaining-rapid-titration-kit-ua079055","title":"Baculovirus Immunostaining Rapid Titration Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eRapid Test Kit for Baculovirus Titer Using Immunostaining\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e\u003cspan\u003eDry ice transportation. Refer to the \"Product Composition\" table for reagent storage.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan\u003eThe rapid assay kit for baculovirus titer using immunostaining specifically recognizes the GP64 protein expressed on the surface of infected cells via a baculovirus GP64 antibody, enabling rapid quantification of baculovirus titers. The entire process takes only 2–3 days, whereas traditional plaque assays and the tissue culture infectious dose 50 (TCID50) method typically require 5–8 days, are labor-intensive, highly dependent on the technician's experience, and involve subjective interpretation of results. Compared with qPCR, which can complete testing within a few hours, qPCR quantifies viral nucleic acids and cannot distinguish between infectious and non-infectious virus particles; in contrast, this product directly detects the GP64 protein, measuring the titer of infectious units (IFU\/mL), yielding results that more closely reflect actual infection efficiency and providing a more reliable basis for precise MOI calculation.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eIn addition, this kit employs a newly developed coating material that not only effectively prevents viral spread through the culture medium but also exhibits significantly lower viscosity compared to traditional methylcellulose. Moreover, it remains liquefied at 37°C, thereby reducing the adverse effects of overheated, overly viscous coatings on cells. The kit is easy to use, simple to remove, and helps minimize human error. With high batch-to-batch precision and excellent reproducibility, this product is suitable for determining the titer of AcMNPV-type baculoviruses, providing rapid and accurate technical support for virus quantification and large-scale production in baculovirus expression systems.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan\u003eThe components and specifications of the baculovirus rapid titer assay kit are as follows. By following the example experimental procedure, you can perform\u003c\/span\u003e\u003cspan\u003e100\u003c\/span\u003e\u003cspan\u003etests.\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSpecifications\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eComponents\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eQuantity\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eStorage temperature\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"5\" style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e100T\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eMouse\u003c\/span\u003e\u003cspan\u003eGP64\u003c\/span\u003e\u003cspan\u003eantibody (\u003c\/span\u003e\u003cspan\u003e1000×\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e2.5\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e-20\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003egoat anti-mouse antibody\u003c\/span\u003e\u003cspan\u003e-HRP\u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003e1000×\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e-20\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003egoat serum (\u003c\/span\u003e\u003cspan\u003e10×\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1.5 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e-20\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003echromogenic substrate\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e4\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003ecoating solution (\u003c\/span\u003e\u003cspan\u003e10×\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0.5 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eRT\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eReagent preparation:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003e4%\u003c\/span\u003e\u003cspan\u003eParaformaldehyde solution: Weigh out\u003c\/span\u003e\u003cspan\u003e4g\u003c\/span\u003e\u003cspan\u003eParaformaldehyde was placed in a glass beaker, and then added…\u003c\/span\u003e\u003cspan\u003e50 mL\u003c\/span\u003e\u003cspan\u003eIn distilled water, use a magnetic stirrer in a fume hood to raise the temperature to\u003c\/span\u003e\u003cspan\u003eSixty\u003c\/span\u003e\u003cspan\u003e℃ (not exceeding\u003c\/span\u003e\u003cspan\u003eSixty-five\u003c\/span\u003e\u003cspan\u003e℃), add while stirring\u003c\/span\u003e\u003cspan\u003eThirty\u003c\/span\u003e\u003cspan\u003e~\u003c\/span\u003e\u003cspan\u003eFifty\u003c\/span\u003e\u003cspan\u003eμL\u003c\/span\u003e\u003cspan\u003eof\/'s\u003c\/span\u003e\u003cspan\u003e4\u003c\/span\u003e\u003cspan\u003eN\u003c\/span\u003e\u003cspan\u003eSodium hydroxide is used to neutralize the solution, with\u003c\/span\u003e\u003cspan\u003epH\u003c\/span\u003e\u003cspan\u003eObtained from a test strip\u003c\/span\u003e\u003cspan\u003epH\u003c\/span\u003e\u003cspan\u003eValue, ultimately\u003c\/span\u003e\u003cspan\u003epH\u003c\/span\u003e\u003cspan\u003eThe value should be\u003c\/span\u003e\u003cspan\u003epH 6\u003c\/span\u003e\u003cspan\u003e~\u003c\/span\u003e\u003cspan\u003e8\u003c\/span\u003e\u003cspan\u003e, once the solution has cooled, add\u003c\/span\u003e\u003cspan\u003e50 mL 0.2 M\u003c\/span\u003e\u003cspan\u003eSodium phosphate solution\u003c\/span\u003e\u003cspan\u003e(pH 7.4)\u003c\/span\u003e\u003cspan\u003eDilution, final\u003c\/span\u003e\u003cspan\u003epH\u003c\/span\u003e\u003cspan\u003eThe value should be\u003c\/span\u003e\u003cspan\u003eseven\u003c\/span\u003e\u003cspan\u003e~\u003c\/span\u003e\u003cspan\u003e8\u003c\/span\u003e\u003cspan\u003e, adjust as necessary. Use\u003c\/span\u003e\u003cspan\u003e0.45 μm\u003c\/span\u003e\u003cspan\u003eMembrane filtration, followed by aliquoting and storage at –\u003c\/span\u003e\u003cspan\u003eTwenty\u003c\/span\u003e\u003cspan\u003e℃, before use from–\u003c\/span\u003e\u003cspan\u003eTwenty\u003c\/span\u003e\u003cspan\u003e℃ Remove and allow to return to room temperature.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003ePBST\u003c\/span\u003e\u003cspan\u003eSolution\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003eWeigh out\u003c\/span\u003e\u003cspan\u003eNa2HPO4 1.42 g\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003eKH2PO4 0.24g\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003eKCl 0.2g\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003eNaCl 8g\u003c\/span\u003e\u003cspan\u003einto the beaker, add\u003c\/span\u003e\u003cspan\u003e800mL\u003c\/span\u003e\u003cspan\u003eultrapure water, and after all has dissolved, add\u003c\/span\u003e\u003cspan\u003e0.5 mL Tween 20\u003c\/span\u003e\u003cspan\u003e, stir well, and make up to volume with ultrapure water to\u003c\/span\u003e\u003cspan\u003e1L\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003e1×\u003c\/span\u003e\u003cspan\u003egoat serum: dilute\u003c\/span\u003e\u003cspan\u003e10×\u003c\/span\u003e\u003cspan\u003egoat serum with\u003c\/span\u003e\u003cspan\u003ePBST\u003c\/span\u003e\u003cspan\u003esolution by a factor of\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003etimes\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan\u003e1×\u003c\/span\u003e\u003cspan\u003emouse\u003c\/span\u003e\u003cspan\u003eGP64\u003c\/span\u003e\u003cspan\u003eantibody: dilute\u003c\/span\u003e\u003cspan\u003e1000×\u003c\/span\u003e\u003cspan\u003emouse\u003c\/span\u003e\u003cspan\u003eGP64\u003c\/span\u003e\u003cspan\u003eantibody with\u003c\/span\u003e\u003cspan\u003e1×\u003c\/span\u003e\u003cspan\u003egoat serum diluted\u003c\/span\u003e\u003cspan\u003eby a factor of\u003c\/span\u003e\u003cspan\u003e1000\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan\u003e1×\u003c\/span\u003e\u003cspan\u003egoat anti-mouse antibody\u003c\/span\u003e\u003cspan\u003e-HRP\u003c\/span\u003e\u003cspan\u003e: dilute\u003c\/span\u003e\u003cspan\u003e1000×\u003c\/span\u003e\u003cspan\u003egoat anti-mouse antibody\u003c\/span\u003e\u003cspan\u003e-HRP\u003c\/span\u003e\u003cspan\u003ewith\u003c\/span\u003e\u003cspan\u003e1×\u003c\/span\u003e\u003cspan\u003egoat serum diluted\u003c\/span\u003e\u003cspan\u003eby a factor of\u003c\/span\u003e\u003cspan\u003e1000\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e2\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e, baculovirus infection:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003ein\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003ewell cell culture plate, one row (\u003c\/span\u003e\u003cspan\u003e12\u003c\/span\u003e\u003cspan\u003ewells) is inoculated with cells in the early logarithmic growth phase\u003c\/span\u003e\u003cspan\u003eSf9\u003c\/span\u003e\u003cspan\u003eCells (per well\u003c\/span\u003e\u003cspan\u003e6.5\u003c\/span\u003e\u003cspan\u003e~\u003c\/span\u003e\u003cspan\u003e7\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e4\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003ecells)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003eIncubate the culture plate at\u003c\/span\u003e\u003cspan\u003e28\u003c\/span\u003e\u003cspan\u003e℃ for\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e\u0026gt;2 hours\u003c\/span\u003e\u003cspan\u003eor overnight\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eTake\u003c\/span\u003e\u003cspan\u003e50 μL\u003c\/span\u003e\u003cspan\u003eof the virus sample and prepare a series of diluted virus samples using culture medium as the diluent. Adjust the dilution range as needed, ensuring thorough mixing between each dilution step. This instruction uses virus sample dilutions of\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e4\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e5\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e6\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003eas examples; this dilution range is suitable for virus samples with titers ranging from\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e6\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003eto\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e9\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eIFU\/mL\u003c\/span\u003e\u003cspan\u003efor the virus sample\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan\u003eCarefully aspirate the culture medium, taking care not to scrape the cells at the bottom of the wells\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan\u003eAdd\u003c\/span\u003e\u003cspan\u003e25\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eof the virus dilution\u003c\/span\u003e\u003cspan\u003esample\u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e4\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003ediluted\u003c\/span\u003e\u003cspan\u003esample\u003c\/span\u003e\u003cspan\u003etake\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003ewells,\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e5\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e6\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003ediluted\u003c\/span\u003e\u003cspan\u003esamples\u003c\/span\u003e\u003cspan\u003eeach take\u003c\/span\u003e\u003cspan\u003e4\u003c\/span\u003e\u003cspan\u003ewells), add\u003c\/span\u003e\u003cspan\u003e25 μL to the negative control wells\u003c\/span\u003e\u003cspan\u003eAdd the culture medium, gently shake the plate to evenly distribute the virus\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6)\u003c\/span\u003e\u003cspan\u003eSeal the culture plate with sealing film,\u003c\/span\u003e\u003cspan\u003e28\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003eIncubate\u003c\/span\u003e\u003cspan\u003e1hr\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7)\u003c\/span\u003e\u003cspan\u003ePrepare in advance\u003c\/span\u003e\u003cspan\u003e15min\u003c\/span\u003e\u003cspan\u003ePut\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003ethe overlay together with\u003c\/span\u003e\u003cspan\u003eSf9\u003c\/span\u003e\u003cspan\u003ecells completely\u003c\/span\u003e\u003cspan\u003ein the culture medium at\u003c\/span\u003e\u003cspan\u003e37\u003c\/span\u003e\u003cspan\u003e℃ water bath to preheat\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8)\u003c\/span\u003e\u003cspan\u003eAspirate the inoculum, avoiding scraping the cells at the bottom of the wells, and dilute the\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003eoverlay layer with\u003c\/span\u003e\u003cspan\u003eSf9\u003c\/span\u003e\u003cspan\u003ecell complete\u003c\/span\u003e\u003cspan\u003eculture medium to\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003eoverlay layer,\u003c\/span\u003e\u003cspan\u003eadd\u003c\/span\u003e\u003cspan\u003e50 μL\u003c\/span\u003e\u003cspan\u003eof\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003eoverlay layer to each well,\u003c\/span\u003e\u003cspan\u003eseal with sealing film, and incubate at\u003c\/span\u003e\u003cspan\u003e28\u003c\/span\u003e\u003cspan\u003e℃ for\u003c\/span\u003e\u003cspan\u003e43\u003c\/span\u003e\u003cspan\u003e-\u003c\/span\u003e\u003cspan\u003e48hr\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e, baculovirus titer detection:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003eadd carefully\u003c\/span\u003e\u003cspan\u003e150\u003c\/span\u003e\u003cspan\u003eμL\u003c\/span\u003e\u003cspan\u003eof\u003c\/span\u003e\u003cspan\u003e4%\u003c\/span\u003e\u003cspan\u003eparaformaldehyde (which needs to be brought back to room temperature in advance\u003c\/span\u003e\u003cspan\u003eand kept at room temperature\u003c\/span\u003e\u003cspan\u003e) into each well containing the overlay, and incubate at room temperature\u003c\/span\u003e\u003cspan\u003efor 30min\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003ediscard the liquid in the wells, and gently tap the\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003ewell plate\u003c\/span\u003e\u003cspan\u003eon a paper towel, then add\u003c\/span\u003e\u003cspan\u003e200\u003c\/span\u003e\u003cspan\u003eμL\u003c\/span\u003e\u003cspan\u003ePBST\u003c\/span\u003e\u003cspan\u003esolution, wash\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003e~\u003c\/span\u003e\u003cspan\u003e4\u003c\/span\u003e\u003cspan\u003etimes, each time incubate on the shaker at low speed for\u003c\/span\u003e\u003cspan\u003e5min\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eAdd to each well\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμL\u003c\/span\u003e\u003cspan\u003eof\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003egoat serum, incubate at room temperature on a shaker at low speed for\u003c\/span\u003e\u003cspan\u003e20min\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan\u003eDiscard the liquid in the wells, place the\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003ewell plate\u003c\/span\u003e\u003cspan\u003egently tap it on paper towels,\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eno washing required\u003c\/span\u003e\u003cspan\u003e, add to each well\u003c\/span\u003e\u003cspan\u003e25\u003c\/span\u003e\u003cspan\u003eμL of 1\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003emouse\u003c\/span\u003e\u003cspan\u003eGP64\u003c\/span\u003e\u003cspan\u003eantibody,\u003c\/span\u003e\u003cspan\u003e37\u003c\/span\u003e\u003cspan\u003e℃ incubation\u003c\/span\u003e\u003cspan\u003e25min\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan\u003eDiscard the liquid in the wells, gently tap the\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003ewell plate\u003c\/span\u003e\u003cspan\u003eon paper towels, and add 200\u003c\/span\u003e\u003cspan\u003eμL\u003c\/span\u003e\u003cspan\u003ePBST\u003c\/span\u003e\u003cspan\u003ePBST\u003c\/span\u003e\u003cspan\u003esolution, wash\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003etimes, each time incubate on the shaker at low speed for\u003c\/span\u003e\u003cspan\u003e5min\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6)\u003c\/span\u003e\u003cspan\u003eAdd to each well\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμL of 1\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan\u003egoat anti-mouse antibody\u003c\/span\u003e\u003cspan\u003e-HRP\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003eincubate at\u003c\/span\u003e\u003cspan\u003e37\u003c\/span\u003e\u003cspan\u003e℃ for\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e25min\u003c\/span\u003e\u003cspan\u003e7)\u003c\/span\u003e\u003cspan\u003eDiscard the liquid in the wells, take the\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003ePat gently with a tissue, then add to each pore.\u003c\/span\u003e\u003cspan\u003e200\u003c\/span\u003e\u003cspan\u003eμL\u003c\/span\u003e\u003cspan\u003ePBST\u003c\/span\u003e\u003cspan\u003eSolution, washing\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003eNext, incubate the plate on the shaker at low speed each time.\u003c\/span\u003e\u003cspan\u003e5 minutes\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8)\u003c\/span\u003e\u003cspan\u003eAdd to each well\u003c\/span\u003e\u003cspan\u003eFifty\u003c\/span\u003e\u003cspan\u003eμL\u003c\/span\u003e\u003cspan\u003eChromogenic substrate (to be prepared in advance)\u003c\/span\u003e\u003cspan\u003eRestore to room temperature\u003c\/span\u003e\u003cspan\u003e), incubate at room temperature\u003c\/span\u003e\u003cspan\u003e1.5-3 hours\u003c\/span\u003e\u003cspan\u003e. After adding the chromogenic substrate, the fastest\u003c\/span\u003e\u003cspan\u003e10 minutes\u003c\/span\u003e\u003cspan\u003eThis allows for a preliminary estimation of the viral titer, though with relatively low accuracy.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e9)\u003c\/span\u003e\u003cspan\u003eInfection foci in each well (blue‑purple cell aggregates) were counted under a microscope.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e10)\u003c\/span\u003e\u003cspan\u003eContaining\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003e~\u003c\/span\u003e\u003cspan\u003e25\u003c\/span\u003e\u003cspan\u003eThe virus titer is calculated based on the dilution of a single infection focus: Virus titer (\u003c\/span\u003e\u003cspan\u003eIFU\/mL\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003cspan\u003e=\u003c\/span\u003e\u003cspan\u003eAverage number of lesions per well × dilution factor ×\u003c\/span\u003e\u003cspan\u003e40\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eIt is not recommended to mix different batches.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"100T","offer_id":43027762675787,"sku":"UA079055-100T","price":900.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_d8b31e29-1edf-4cd9-ad6c-97210d6a5587.png?v=1789106460"},{"product_id":"high-fidelity-dna-polymerase-kit-ua070141","title":"High-Fidelity DNA Polymerase Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e\u003cspan\u003eStore at -25 ~ -15℃ for 2 years\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cdiv align=\"center\"\u003e\u003cfigure class=\"table\"\u003e\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" style=\"border-collapse:collapse;border:none;\" width=\"461\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:167.65pt;\" width=\"224\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border-left:none;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:92.15pt;\" width=\"123\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e100 Rxns\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border-left:none;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:86.2pt;\" width=\"115\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e250 Rxns\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border-top:none;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:167.65pt;\" width=\"224\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003eHigh-Fidelity DNA polymerase\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:92.15pt;\" width=\"123\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e200 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:86.2pt;\" width=\"115\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e500 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border-top:none;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:167.65pt;\" width=\"224\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003eHigh-Fidelity Reaction buffer (5×)\u003csup\u003ea\u003c\/sup\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:92.15pt;\" width=\"123\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e1 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:86.2pt;\" width=\"115\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e2.5 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border-top:none;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:167.65pt;\" width=\"224\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003edNTP mix (10 mM)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:92.15pt;\" width=\"123\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e150 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:86.2pt;\" width=\"115\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e400 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border-top:none;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:167.65pt;\" width=\"224\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003eRNase Free\u003c\/span\u003e\u003cspan style=\"color:black;\"\u003e\u003cspan lang=\"EN-US\"\u003e \u003c\/span\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003edH\u003csub\u003e2\u003c\/sub\u003eO\u003c\/span\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:92.15pt;\" width=\"123\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e2×1.5mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:86.2pt;\" width=\"115\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e5×1.5mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border-top:none;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:167.65pt;\" width=\"224\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003eMgCl\u003csub\u003e2\u003c\/sub\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:92.15pt;\" width=\"123\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e500μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:86.2pt;\" width=\"115\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e1 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border-top:none;border:solid windowtext 1.0pt;padding:0cm 5.4pt 0cm 5.4pt;width:167.65pt;\" width=\"224\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003eDMSO\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:92.15pt;\" width=\"123\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e200 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border-bottom:solid windowtext 1.0pt;border-left:none;border-right:solid windowtext 1.0pt;border-top:none;padding:0cm 5.4pt 0cm 5.4pt;width:86.2pt;\" width=\"115\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e600 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/figure\u003e\u003c\/div\u003e\n\u003cp\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003ea. High-Fidelity Reaction buffer (5×)\u003c\/span\u003e\u003cspan style=\"background-color:#FBFBFB;color:#080808;\"\u003e\u003cspan style=\"letter-spacing:.2pt;\"\u003econtains\u003c\/span\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;letter-spacing:.2pt;'\u003e1\u003c\/span\u003e\u003cspan class=\"katex-mathml\" lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;letter-spacing:.2pt;'\u003e0 mM\u003c\/span\u003e\u003cspan style=\"letter-spacing:.2pt;\"\u003eof\u003c\/span\u003e\u003cspan lang=\"EN-US\" style=\"letter-spacing:.2pt;\"\u003e \u003c\/span\u003e\u003cspan class=\"katex-mathml\" lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;letter-spacing:.2pt;'\u003eMg\u003csup\u003e2+\u003c\/sup\u003e\u003c\/span\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;letter-spacing:.2pt;'\u003e,\u003c\/span\u003e\u003cspan style=\"letter-spacing:.2pt;\"\u003eand in the\u003c\/span\u003e\u003cspan class=\"katex-mathml\" lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;letter-spacing:.2pt;'\u003e1\u003c\/span\u003e\u003c\/span\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e×\u003c\/span\u003e\u003cspan style=\"background-color:#FBFBFB;color:#080808;\"\u003e\u003cspan style=\"letter-spacing:.2pt;\"\u003eworking solution, the final concentration is\u003c\/span\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;letter-spacing:.2pt;'\u003e2 mM\u003c\/span\u003e\u003cspan style=\"letter-spacing:.2pt;\"\u003e, so no additional addition is required for routine amplification; the separately provided\u003c\/span\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;letter-spacing:.2pt;'\u003e25mM\u003c\/span\u003e\u003c\/span\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003eMgCl\u003csub\u003e2\u003c\/sub\u003e\u003c\/span\u003e\u003cspan\u003eis only used for optimizing conditions in specific primer\u003c\/span\u003e\u003cspan lang=\"EN-US\" style='font-family:\"Times New Roman\",serif;'\u003e\/\u003c\/span\u003e\u003cspan\u003etemplate systems.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e1.\u003c\/span\u003e\u003cstrong\u003eSystem Preparation\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eAll operations should be performed on ice throughout the process. After thawing, thoroughly mix each component and promptly return it to -20°C storage after use.\u003c\/span\u003e\u003c\/p\u003e\n\u003cfigure class=\"table\"\u003e\u003ctable style=\"border-collapse:collapse;border:1px solid #000000;width:auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eComponents\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e25\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµL system\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eFinal Concentration\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eTemplate DNA\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eX\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eN\/A\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eHigh-Fidelity DNA polymerase\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1U\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eForward Primer (10\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµM\u003c\/span\u003e\u003cspan style=\"color:#000000;\"\u003e)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e0.75\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e0.3\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eReverse Primer (10\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµM\u003c\/span\u003e\u003cspan style=\"color:#000000;\"\u003e)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e0.75\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e0.3\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003e10mM dNTP mix\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e0.75\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e0.3 m\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eM\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eHigh-Fidelity Reaction Buffer (5\u003c\/span\u003e\u003cspan\u003e×\u003c\/span\u003e\u003cspan style=\"color:#000000;\"\u003e)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1 ×\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eRNase-Free\u003c\/span\u003e\u003cspan style=\"color:#000000;\"\u003edH\u003csub\u003e2\u003c\/sub\u003eO\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eAdjust to 25\u003c\/span\u003e\u003cspan style=\"color:#080808;\"\u003eµL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eN\/A\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003e25mM MgCl\u003csub\u003e2\u003c\/sub\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eOptional\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eN\/A\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/figure\u003e\u003cp\u003e\u003cbr\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003ea. The reaction volume can be adjusted between 10 and 50 μL. For volumes other than 25 μL, the reagents should be diluted proportionally. Reaction volumes greater than 50 μL are not recommended;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eb. For most reactions, Mg\u003csup\u003e2+\u003c\/sup\u003ethe optimal final concentration is 1.5–2 mM. The system already contains a final concentration of 2 mM Mg\u003csup\u003e2+\u003c\/sup\u003e; if necessary, the 25 mM MgCl\u003csub\u003e2\u003c\/sub\u003eprovided in the kit can be used to incrementally adjust the Mg\u003csup\u003e2+\u003c\/sup\u003econcentration in steps of 0.2–0.5 mM to determine the optimal concentration;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003ec. After preparation, gently tap the bottom of the tube to mix thoroughly and briefly centrifuge; avoid vigorous vortexing;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003ed. The optimal reaction concentration varies depending on the template; the following table shows the recommended template amounts for a 25 μL reaction:\u003c\/span\u003e\u003c\/p\u003e\n\u003cfigure class=\"table\"\u003e\u003ctable style=\"border-collapse:collapse;border:1px solid #000000;width:auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eTemplate\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e\u0026lt;1 kb\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1–10 kb\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e\u0026gt;10 kb\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003ePlasmid or viral DNA\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e50 pg–50 ng\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e50 pg–50 ng\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e5 ng–50 ng\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003ecDNA\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd colspan=\"3\" style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e0.5 – 2.5 μL (not exceeding 1\/10 of the total PCR reaction volume)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/figure\u003e\u003cp\u003e\u003cbr\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e2.\u003c\/span\u003e\u003cstrong\u003eReaction program\u003c\/strong\u003e\u003c\/p\u003e\n\u003cfigure class=\"table\"\u003e\u003ctable style=\"border-collapse:collapse;border:1px solid #000000;width:auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eCycling steps\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eTemperature\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eTime\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eNumber of cycles\u003csup\u003ed\u003c\/sup\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003ePre-denaturation\u003csup\u003ea\u003c\/sup\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e95℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e2 min\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eDenaturation\u003csup\u003eb\u003c\/sup\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e98℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e20 sec\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd rowspan=\"3\" style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\n\u003cp style=\"text-align:center;\"\u003e \u003c\/p\u003e\n\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e25-35 cycles\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eAnnealing\u003csup\u003ec\u003c\/sup\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e58-72℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e10-15 sec\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eExtension\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e68\/72℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e30-60 sec\/kb\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eFinal extension\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e72℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e5-10 min\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"background-color:#E7E6E6;border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#000000;\"\u003eHold\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e12℃\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan style=\"color:#080808;\"\u003e∞\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e-\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/figure\u003e\u003cp\u003e\u003cbr\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003ea. Pre-denaturation conditions: For simple plasmids\/λDNA, 95℃ for 2 minutes is sufficient; for complex genomic DNA, 95℃ for 3–5 minutes is recommended.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eb. Denaturation conditions: 98℃ for 20 seconds is recommended. If the fragment is \u0026gt;10 kb, the denaturation temperature can be lowered to 92–94℃ for 15 seconds to reduce thermal damage.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003ec. Annealing temperature: In the High-Fidelity Reaction buffer system, primer annealing kinetics may be slightly altered under high-fidelity optimization. It is recommended to set the annealing time to 10–15 seconds. For initial experiments, a temperature gradient is recommended to determine the optimal annealing temperature.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003ed. Number of cycles: To maintain the highest fidelity, it is advised to limit the number of cycles to ≤25–35 cycles.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e3.\u003c\/span\u003e\u003cstrong\u003ePCR procedure for high GC content and difficult templates\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eWhen the GC content of the template amplicon exceeds 60% or when the template has complex secondary structures, it is recommended to adjust the protocol as follows: on the basis of the standard reaction system, introduce a solvent additive—add DMSO—at a final concentration of 2%–8% in a 50 μL reaction (i.e., add 1–4 μL; it is recommended to explore using increments of 1% as a gradient).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e(Note: Excessively high concentrations of the cosolvent may slightly reduce enzyme fidelity; it is recommended to use it only for challenging templates.)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003ea. Ice-bench operations: High-Fidelity DNA Polymerase is a hot-start enzyme system; although it allows for room-temperature reaction setup, performing operations on ice can yield exceptionally uniform experimental results.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"100Rxns","offer_id":43195884732491,"sku":"UA070141-100Rxns","price":100.0,"currency_code":"USD","in_stock":true},{"title":"250Rxns","offer_id":43195884765259,"sku":"UA070141-250Rxns","price":200.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/75b4836e34c94c01bb48e32b08643c29_12ddefbf-19c7-41ee-a199-d0413d650bac.png?v=1789621344"},{"product_id":"lipo-nb-transfection-reagents-ua079058","title":"Lipo-UA Transfection Reagents","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e\u003cspan\u003e4℃, expiration date see reagent bottle label\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cspan\u003eCell transfection reagents facilitate the entry of nucleic acids into cells and enable the expression of the target gene within the cells. Liposomes are among the most commonly used and highly efficient transfection reagents in laboratories. Lipo-UA liposome cell transfection reagent is a highly effective DNA transfection reagent, offering advantages such as high transfection efficiency, broad applicability, ease of use, and cost-effectiveness, making it suitable for DNA transfection experiments in most cell types.\u003c\/span\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:justify;\"\u003e\u003cspan\u003eThe liposome cell transfection kit consists of liposome reagent L and transfection enhancer E, with the following specifications. The number of 96-well plates that can be transfected is calculated based on a transfection of 0.1 μg DNA per well, and the ratio of transfection reagents DNA, L, and E is calculated as DNA (μg) : E (μL) : L (μL) = 1:2:3.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify;\"\u003e \u003c\/p\u003e\n\u003cfigure class=\"table\"\u003e\u003ctable style=\"border-collapse:collapse;border:1px solid #000000;width:auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cstrong\u003eSpecifications\u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cstrong\u003eLiposome Reagent L\u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cstrong\u003eTransfection Enhancer E\u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cstrong\u003eNumber of 96-well plates that can be transfected\u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cstrong\u003eAmount of DNA that can be transfected (μg)\u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1.5 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1.5 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e50\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e500\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e5 x 1.5 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e5 x 1.5 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e5 x 1 ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e250\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003e2500\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/figure\u003e\u003cp\u003e\u003cbr\u003e \u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003eAt\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003eConfucius\u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003eor other cell culture plates of various sizes\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003cspan\u003eSeed the cells to be transfected at an appropriate density, per well.\u003c\/span\u003e\u003cspan\u003e100\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003eCell,\u003c\/span\u003e\u003cspan\u003eIncubate overnight.\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eCell density during transfection\u003c\/span\u003e\u003cspan\u003etransfection efficiency\u003c\/span\u003e\u003cspan\u003e\/\u003c\/span\u003e\u003cspan\u003eCytotoxicity has a significant impact, and different cell lines require optimization; typically, for example,\u003c\/span\u003e\u003cspan\u003eHEK293\u003c\/span\u003e\u003cspan\u003eor\u003c\/span\u003e\u003cspan\u003eHeLa\u003c\/span\u003e\u003cspan\u003eCell recommendation\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eDuring transfection\u003c\/span\u003e\u003cspan\u003eThe cell density is at\u003c\/span\u003e\u003cspan\u003e80%\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eleft and right\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eCalculate the required amount based on the area of the transfected cells.\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eTotal amount, transfection enhancer\u003c\/span\u003e\u003cspan\u003eE\u003c\/span\u003e\u003cspan\u003eand liposome reagents\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eTotal reagent volume. Recommended.\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eDosage:\u003c\/span\u003e\u003cspan\u003e6\u003c\/span\u003e\u003cspan\u003eOrifice plate, per hole\u003c\/span\u003e\u003cspan\u003e2.5\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg DNA\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003e24\u003c\/span\u003e\u003cspan\u003eOrifice plate, per hole\u003c\/span\u003e\u003cspan\u003e0.5\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg DNA\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003eOrifice plate, per hole\u003c\/span\u003e\u003cspan\u003e0.1\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg DNA\u003c\/span\u003e\u003cspan\u003e. Recommended\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003eE\u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eReagent ratio as per\u003c\/span\u003e\u003cspan\u003eDNA (\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg)\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003eE (\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eL)\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003eL (\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eL)\u003c\/span\u003e\u003cspan\u003eFor\u003c\/span\u003e\u003cspan\u003e1:2:3\u003c\/span\u003e\u003cspan\u003ePrepare the transfection reagent. If significant cytotoxicity is observed, reduce the amount accordingly.\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eReagent\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\u003c\/span\u003e\u003cspan\u003eTransfection reagent preparation: The following is for transfecting one plate.\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003eHole experiment board\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan style=\"color:#EE0000\"\u003e \u003c\/span\u003e\u003cspan\u003ePer hole\u003c\/span\u003e\u003cspan\u003e0.1\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg DNA\u003c\/span\u003e\u003cspan\u003eConfigure as an example\u003c\/span\u003e\u003cspan\u003e1 mL\u003c\/span\u003e\u003cspan\u003eTransfection reagent. Transfection\u003c\/span\u003e\u003cspan\u003eOther cell culture plates of various sizes are according to\u003c\/span\u003e\u003cspan\u003eDNA\u003c\/span\u003e\u003cspan\u003eAdjust the amount of transfection reagent added to the culture medium. For example,\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003eHole experiment board\u003c\/span\u003e\u003cspan\u003e100\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eThe culture medium is supplemented with the following prepared components:\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eTransfection reagent\u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003ePer hole\u003c\/span\u003e\u003cspan\u003e0.1\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg DNA)\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e24\u003c\/span\u003e\u003cspan\u003ePer well of the experimental plate\u003c\/span\u003e\u003cspan\u003e500\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eThe culture medium is supplemented with the following prepared components:\u003c\/span\u003e\u003cspan\u003eFifty\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eTransfection reagent\u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003ePer hole\u003c\/span\u003e\u003cspan\u003e0.5\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg DNA)\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003e6\u003c\/span\u003e\u003cspan\u003ePer well of the experimental plate\u003c\/span\u003e\u003cspan\u003e2500\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eThe culture medium is supplemented with the following prepared components:\u003c\/span\u003e\u003cspan\u003e250\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eTransfection reagent\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003ePer hole\u003c\/span\u003e\u003cspan\u003e2.5\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg DNA)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003ea)\u003c\/span\u003e\u003cspan\u003euse\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eOpti MEM I\u003c\/span\u003e\u003cspan\u003eDilute the nucleic acid to\u003c\/span\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003emu\u003c\/span\u003e\u003cspan\u003eg\/mL\u003c\/span\u003e\u003cspan\u003e, take\u003c\/span\u003e\u003cspan\u003e500\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL,\u003c\/span\u003e\u003cspan\u003eadd\u003c\/span\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003etransfection enhancer\u003c\/span\u003e\u003cspan\u003eE\u003c\/span\u003e\u003cspan\u003e, mix well and incubate at room temperature\u003c\/span\u003e\u003cspan\u003e5 min\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003eb)\u003c\/span\u003e\u003cspan\u003etake\u003c\/span\u003e\u003cspan\u003e500\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL Opti MEM I\u003c\/span\u003e\u003cspan\u003e, add\u003c\/span\u003e\u003cspan\u003e30\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eliposome reagent\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003e, mix well\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003ec)\u003c\/span\u003e\u003cspan\u003eadd all of the liposome reagent\u003c\/span\u003e\u003cspan\u003eb\u003c\/span\u003e\u003cspan\u003eprepared in step\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eto the nucleic acid and transfection enhancer\u003c\/span\u003e\u003cspan\u003ea\u003c\/span\u003e\u003cspan\u003emixture prepared in step\u003c\/span\u003e\u003cspan\u003eE\u003c\/span\u003e\u003cspan\u003e, mix well, and incubate at room temperature\u003c\/span\u003e\u003cspan\u003efor 20 min\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4.\u003c\/span\u003e\u003cspan\u003edrop the transfection reagent prepared in step\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003einto the corresponding cell culture wells, adding\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eper well, and gently shake a few times to mix\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5.\u003c\/span\u003e\u003cspan\u003eaccording to experimental needs, after transfection for a certain period of time, such as\u003c\/span\u003e\u003cspan\u003e24 hr–96 hr\u003c\/span\u003e\u003cspan\u003edetect the expression of the target gene\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eIt is not recommended to mix different batches.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"1.5ml","offer_id":48960960004171,"sku":"UA079058-1.5ml","price":125.0,"currency_code":"USD","in_stock":true},{"title":"5*1.5ml","offer_id":48960960036939,"sku":"UA079058-5*1.5ml","price":490.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/3407a553-1893-4160-a4aa-e4f856801c37_24a1dde5-2030-413e-8db2-df7073bd1e13.png?v=1790769699"}],"url":"https:\/\/www.antbioinc.com\/collections\/other-kit.oembed?page=3","provider":"AntBio","version":"1.0","type":"link"}