25 Hydroxy Vitamin D BioAssay(TM) ELISA Kit (Human)

Product Overview

• Article Name: 25 Hydroxy Vitamin D BioAssay(TM) ELISA Kit (Human)
• Biozol Catalog Number: USB-369682
• Supplier Catalog Number: 369682
• Alternative Catalog Number: USB-369682-48,USB-369682-96
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

The 25 Hydroxy Vitamin D BioAssay(TM) ELISA Kit is a quantitative competitive assay for the detection of 25 Hydroxy Vitamin D in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples. Detection Range: 3.125-200ng/ml Sensitivity: 1.875ng/ml Precision: Intra-assay CV: <8% Inter-assay CV: <10% Assay Principle: The microtiter plate provided in this kit has been pre-coated with 25 Hydroxy Vitamin D. 25 Hydroxy Vitamin D in the standard or sample competes with a fixed amount of plate-coated 25 Hydroxy Vitamin D for antibody binding sites on the Antibody-Biotin reagent. Excess conjugate and unbound sample or standard are washed from the plate, after which Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. TMB substrate is then added to each well. The enzyme-substrate reaction is terminated by the addition of a sulfuric acid solution and the resulting color is measured spectrophotometrically at 450nm. The concentration of HS in the samples is determined by an inverse correlation between 25 Hydroxy Vitamin D in the sample and the assay signal intensity. Kit Components: *369682A: Microtiter Strips, 1x48 wells or 1x96 wells *369682B: Standard, 1x1 vial or 2x1 vials 369682C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml 369682D: Antibody (Biotin) (Concentrated), 1x1 vial 369682E: Antibody Dilution Buffer, 1x5ml or 1x10ml 369682F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul 369682G: SABC Dilution Buffer, 1x5ml or 1x10ml 369682H: TMB Substrate, 1x5ml or 1x10ml 369682J: Stop Solution, 1x5ml or 1x10ml 369682K: Wash Buffer, 25X, 1x15ml or 1x30ml 369682L: Purified Water, 1x200ul Storage and Stability: Store unopened *369682A at 4C, store at -20C once opened. Store unopened *369682B at 4C, once reconstituted, store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. 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 and samples to wells! 2. Add 50ul standard or sample to each well. 3. Add 50ul Antibody (Biotin) working solution to each well and incubate for 45 minutes at 37C 4. Aspirate and wash 3 times. 5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37C 6. Aspirate and wash 5 times. 7. Add 90ul TMB substrate. Incubate 10-20 minutes at 37C 8. Add 50ul Stop Solution. Read at 450nm immediately. 9. Calculate results