{"product_id":"human-cyclin-d1-elisa-kit","title":"Human Cyclin D1 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 freeze-thawed.\u003cbr\u003eFinally, centrifuge the homogenate at 5000×g for 5-10 minutes and remove the supernatant for analysis.\u003cbr\u003eCell culture supernatant: Centrifuge at 1000×g for 20 minutes.\u003cbr\u003eRemove the supernatant for analysis or store at -20°C or -80°C, but avoid repeated freeze-thaw cycles.\u003cbr\u003e\u003cbr\u003ePre-Assay Preparation:\u003cbr\u003e1. Remove the reagent 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 stand for 15 minutes to completely dissolve, then gently mix (concentration 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 seven EP tubes and add 500uL of universal diluent to each tube.\u003cbr\u003ePipette 500uL of the 20ng\/mL standard working solution into the first EP tube and mix thoroughly to make a 10ng\/mL standard working solution.\u003cbr\u003eRepeat this procedure for subsequent tubes.\u003cbr\u003eThe last tube is used as a blank well; there is no need to pipette liquid from the penultimate tube.\u003cbr\u003eSee the figure below for details.\u003cbr\u003e3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use.\u003cbr\u003eDilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent).\u003cbr\u003ePrepare and use immediately.\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.\u003cbr\u003eAllow 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.\u003cbr\u003eThis will reduce the impact of matrix effects on the test results.\u003cbr\u003eThe sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration.\u003cbr\u003eIt 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\/8dd110a9cee94ea1a54c8bf38a044a5c.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 cyclin D1 capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of cyclin D1 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 Cyclin D1 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\n\u003cstrong\u003e9\u003c\/strong\u003e \u003cstrong\u003e6\u003c\/strong\u003e \u003cstrong\u003eT\u003c\/strong\u003e \u003cstrong\u003e match \u003c\/strong\u003e \u003cstrong\u003e set \u003c\/strong\u003e\n\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-coating 96 Well plate \u003c\/td\u003e\n\u003ctd style=\"height: 10px; width: 19%; text-align: center;\" width=\"19.0000%\"\u003e8 Hole ×12 Strip \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  branch \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 liquid \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 Bottom   thing   （ TMB ） \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 liquid \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 Zhang \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  Share \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\u003eCyclin D1 is a protein encoded by the CCND1 gene. The CCND1 gene encodes the cyclin D1 protein. This gene is located on the long arm of chromosome 11 (band 11q13). It is 13,388 base pairs long and translates to 295 amino acids. Cyclin D1 is expressed in all adult tissues, except cells derived from bone marrow stem cells (both lymphoid and myeloid lineages). Its overexpression has been shown to be associated with early cancer onset and tumor progression, and it can contribute to tumorigenesis by increasing anchorage-dependent growth and angiogenesis through VEGF production.\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\u003e0.312-20 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 homogenate, cell culture supernatant\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":48367993749749,"sku":null,"price":294.6,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0789\/8274\/2261\/files\/AntBioImage_06a4c5c5-b2f5-4c5e-9025-141aa5c1265d.png?v=1783240855","url":"https:\/\/nortrano.com\/products\/human-cyclin-d1-elisa-kit","provider":"nortrano","version":"1.0","type":"link"}