{"product_id":"human-ppar","title":"Human PPAR-α 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\u003eRequired experimental equipment:\u003cbr\u003e1. Microplate reader (450nm)\u003cbr\u003e2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL\u003cbr\u003e3. 37°C incubator\u003cbr\u003e4. Distilled or deionized water\u003cbr\u003e\u003cbr\u003eSample preparation and requirements:\u003cbr\u003eTissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results).\u003cbr\u003eWeigh and mince the tissue.\u003cbr\u003eAdd the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice.\u003cbr\u003eTo further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed.\u003cbr\u003eFinally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis.\u003cbr\u003e\u003cbr\u003eCell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes.\u003cbr\u003eSuspension cells can be collected directly by centrifugation.\u003cbr\u003eWash the collected cells three times with pre-chilled PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately).\u003cbr\u003eDisrupt the cells by repeated freezing and thawing or sonication.\u003cbr\u003eCentrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis.\u003cbr\u003e\u003cbr\u003eCell Culture Supernatant: Centrifuge at 1000×g for 20 minutes.\u003cbr\u003eCollect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing.\u003cbr\u003e\u003cbr\u003eOther biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test.\u003cbr\u003e\u003cbr\u003ePre-test preparation:\u003cbr\u003e1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature.\u003cbr\u003e2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 20 ng\/mL).\u003cbr\u003eThen dilute to the following concentrations: 20 ng\/mL, 10 ng\/mL, 5 ng\/mL, 2.5 ng\/mL, 1.25 ng\/mL, 0.625 ng\/mL, 0.3125 ng\/mL, and 0 ng\/mL.\u003cbr\u003eSerial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube.\u003cbr\u003ePipette 500 μL of the 20 ng\/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng\/mL standard working solution.\u003cbr\u003eRepeat this procedure for subsequent tubes.\u003cbr\u003eThe last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube.\u003cbr\u003eSee the figure below for details.\u003cbr\u003e\u003cbr\u003e\u003cdiv\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20250221\/c5275922094b48e3bc3f35f743d68dd5.png\" alt=\"\" width=\"468\" height=\"180\"\u003e\u003c\/div\u003e\n\u003cbr\u003e\u003cbr\u003e3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute.\u003cbr\u003eDilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent).\u003cbr\u003ePrepare immediately before use.\u003cbr\u003e4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute.\u003cbr\u003eDilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent).\u003cbr\u003ePrepare immediately.\u003cbr\u003e5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing).\u003cbr\u003e\u003cbr\u003eProcedure:\u003cbr\u003e1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes.\u003cbr\u003eSeal the remaining strips in a ziplock bag and return to 4°C.\u003cbr\u003e2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells.\u003cbr\u003eAdd 100 μL of universal diluent to the blank wells.\u003cbr\u003eCover with a film and incubate at 37°C for 60 minutes.\u003cbr\u003e(Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.)\u003cbr\u003e3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing.\u003cbr\u003eAdd 100 μL of Biotinylated Antibody Working Solution directly to each well.\u003cbr\u003eCover with a film and incubate at 37°C for 60 minutes.\u003cbr\u003e4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well.\u003cbr\u003eLet stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper.\u003cbr\u003eRepeat this process three times (a plate washer can also be used).\u003cbr\u003e5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well.\u003cbr\u003eCover with a film and incubate at 37°C for 30 minutes.\u003cbr\u003e6. Washing: Discard the liquid and wash the plate five times as in step 4.\u003cbr\u003e7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes.\u003cbr\u003e8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well.\u003cbr\u003eImmediately measure the OD value of each well at a wavelength of 450 nm.\u003cbr\u003e\u003cbr\u003eCalculating experimental results:\u003cbr\u003e1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor.\u003cbr\u003ePlot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis.\u003cbr\u003e2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest.\u003cbr\u003eMultiply the sample concentration by the corresponding dilution factor.\u003cbr\u003e\u003cbr\u003e\u003cimg style=\"page-break-inside: avoid; display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin_new_bak-china\/20250312\/350c793c645440d69972fa6f35a84d26.png\" alt=\"\" width=\"512\" height=\"356\"\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 a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against peroxisome proliferator-activated receptor alpha (PPAR-α). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of peroxidase (HRP) and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of peroxisome proliferator-activated receptor alpha (PPAR-α) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSource\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eHuman\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eHuman Peroxisome Proliferator Activated Receptor Alpha ELISA Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eDouble antibody sandwich method\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: 60%; height: 110px; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 10px;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e\u003cstrong\u003e Name \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e\u003cstrong\u003e96-Well Format Configuration \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cstrong\u003e remark \u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e Pre-coated 96-Well Plate \u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e8-Well × 12 Strips\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e without \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e Standard \u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e2  Vials \u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cdiv style=\"text-align: center;\"\u003e Dilute as per instructions \u003c\/div\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px; text-align: center;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e\u003cdiv\u003e Universal diluent \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e\u003cdiv\u003e2×20mL\u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cdiv\u003e without \u003c\/div\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px; text-align: center;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e\u003cdiv\u003e Concentrated biotinylated antibody ( 100× ）  \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e\u003cdiv\u003e120uL\u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cdiv\u003e Dilute as per instructions \u003c\/div\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 31.98%; text-align: center;\"\u003e\u003cdiv\u003e Concentrated enzyme conjugate ( 100× ） \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 19%; text-align: center;\"\u003e\u003cdiv\u003e120uL\u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 29.98%; text-align: center;\"\u003e\u003cdiv\u003e Dilute as per instructions \u003c\/div\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px; text-align: center;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e\u003cdiv\u003e20× Washing Buffer \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e\u003cdiv\u003e2×10mL\u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cdiv\u003e Dilute as per instructions \u003c\/div\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px; text-align: center;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e\u003cdiv\u003e TMB Substrate Solution\u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e\u003cdiv\u003e10mL\u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cdiv\u003e without \u003c\/div\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px; text-align: center;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e\u003cdiv\u003e Stop Solution \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e\u003cdiv\u003e6mL\u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cdiv\u003e without \u003c\/div\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px; text-align: center;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e\u003cdiv\u003e Sealing film \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e\u003cdiv\u003e4 Sheets \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cdiv\u003e without \u003c\/div\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 10px; text-align: center;\"\u003e\n\u003ctd style=\"height: 10px; width: 31.98%; text-align: center;\" width=\"31.9800%\"\u003e\u003cdiv\u003e Instructions \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e\u003cdiv\u003e1 \u003c\/div\u003e\u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 29.98%; text-align: center;\" width=\"29.9800%\"\u003e\u003cdiv\u003e without \u003c\/div\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\u003ePeroxisome proliferator-activated receptor α (PPAR-α), also known as NR1C1 (nuclear receptor subfamily 1, group C, member 1), is a nuclear receptor protein encoded by the PPARA gene. Along with peroxisome proliferator-activated receptor δ and peroxisome proliferator-activated receptor γ, it belongs to the peroxisome proliferator-activated receptor subfamily. It was the first member of the PPAR family cloned by Stephen Green in 1990 and was identified as a nuclear receptor for a distinct class of rodent hepatocarcinogens that promotes peroxisome proliferation. It is primarily activated by ligand binding.\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. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. \u003cbr\u003e2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. \u003cbr\u003e3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. \u003cbr\u003e4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. \u003cbr\u003e5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. \u003cbr\u003e6. Avoid direct exposure to strong light during storage and incubation. \u003cbr\u003e7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. \u003cbr\u003e8. Do not use expired products, and do not mix components with different product numbers and batches. \u003cbr\u003e9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. \u003cbr\u003e10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eIf the unopened kit is stored at 4°C, the shelf life is 6 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\u003e0.31-20 ng\/mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue homogenates, cell lysates, cell culture supernatants, and other biological fluids\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":43216128901195,"sku":"abs5510201-96T","price":368.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/www.antbioinc.com\/products\/human-ppar","provider":"AntBio","version":"1.0","type":"link"}