he Vitamin D3 Receptor (VDR) ELISA Kit (Human) is a sandwich assay for the quantitative detection of VDR in human serum,plasma,tissue homogenates and other biological fluids.Detection Range:0.625-40ng/mlSensitivity:<0.375ng/mlAssay Principle:The microtiter plate provided in this kit has been pre-coated with an antibody specific to VDR. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to VDR. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added,only those wells that contain VDR,Biotin-conjugated antibody and enzyme-conjugated Streptavidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of VDR in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components:*359610A: Microtiter Strips,1x96 wells *359610B: Standard,2x1vial359610C: Sample/Standard Dilution Buffer,1x20ml359610D: Antibody (Biotin) Concentrate,1x120ul359610E: Antibody Dilution Buffer,1x10ml359610F: Streptavidin-HRP Conjugate (SABC),1x120ul359610G: SABC Dilution Buffer,1x10ml359610H: TMB Substrate,1x10ml359610J: Stop Solution,1x10ml359610K: Wash Buffer (25X),1x30mlStorage and Stability:Store unopened *359610A at 4ºC; store at -20ºC once opened. Store unopened *359610B at 4°C; once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product,centrifuge the original vials after thawing and prior to removing the cap.Assay Summary:1. Wash plate 2 times before adding standards,samples and control (zero) to wells!2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 minutes at 37°C4. Aspirate and wash 3 times.5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate and wash 5 times.7. Add 90ul TMB substrate. Incubate 15-30 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately.9. Calculate results.