BSA ELISA Kit
BSA ELISA Kit
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$750 USD
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Product Specification
| Usage |
1. Preparation work: 1 Kit preparation: Each component of the kit is placed at room temperature for equilibrium 30min After starting the operation. 2 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. 3 Reagent preparation: ( Note: 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) ① 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. ( Note: Buffer （ 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) ② Preparation of color developing solution: The color developing 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, generally before use 10min Formulation. Do not use if the colored solution has turned blue after mixing) ③ 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× ）。（ Note: 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) ④ Preparation of standards ( Note: 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:  2. Operation steps: All operations are performed at room temperature and repeat assays are recommended for all spiked wells. 1 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. ( Note: The slats are easy to fall off during the washing process, so be careful to mark them well.) 2 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 。（ Note: 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.) 3 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. ( Note: 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 。） 4 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 。 5 Termination: adding termination solution, 100µL/ Holes, can be read after the color is evenly developed ( Note: Generally add stop solution 20min Complete readings within). 6 Reading: Put the microplate into the microplate reader and set the wavelength to dual wavelengths 450/630nm , read the absorbance value ( Note: It is recommended that it be set in the microplate reader reading program 5-10s Of shock). 3. Data processing: 1 Calculation of absorbance correction value The absorbance correction value of each standard or sample is: OD450nm-OD630nm- Blank control well absorbance values 2 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: Y=((A-D)/(1+(X/C)^B))+D Substitute the sample absorbance correction value into the formula to calculate the sample BSA Content of. (Note the dilution factor)3 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. Note: ① Per Chinese Pharmacopoeia <9012> 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. Example showcase: (The following standard curves are for reference only, and the standard curves drawn in each experiment shall prevail)  |
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| Synonym | Bovine Serum Albumin Residue Detection Kit | ||||||||||||||||||
| Description |
Product background: 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. 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. Reaction principle: 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. Analytical Method Validation Guidance: 1 According to the guidelines of Chinese Pharmacopoeia <9101> 、 <9012> 、 <9401> as well as ICH 2 , 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. 3 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. 4 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. Product performance indicators: 1 Limit of detection: < 0.5ng/mL 2 Limit of quantification: 0.5ng/mL 3 Linear range: 0.5-32ng/mL 4 Accuracy (spiked recovery): 70%-130% 5 Accuracy (measurement deviation): ≤15% 6 , Repeatability (intra-batch difference): ≤10% Specificity: 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.
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| Composition |
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| General Notes | 1. 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. 2. Containers and consumables for measuring BSA content should be dedicated to prevent BSA contamination in the laboratory. 3. The pipette is dedicated or uses a gun tip with a filter element. 4. 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. 5. All components in the kit must be restored to room temperature (20-25 ℃) before use. 6. 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 ℃. 7. 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. 8. 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. 9. 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. 10. 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. 11. When reading, pay attention to checking whether the detection wavelength and fitting equation are correct. 12. 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. 13. 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. 14. 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. 15. This kit is only for in vitro research, not for clinical diagnosis. |
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| Application | The 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. | ||||||||||||||||||
| Storage Temp. | The 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. |
