The Embryonic Growth/Differentiation Factor 1 (GDF1) BioAssay(TM) ELISA Kit is a quantitative sandwich assay for the detection of GDF1 in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples. Detection Range: 31.25-2000pg/ml Sensitivity: 18.75pg/ml Precision: Intra-Assay CV: <8% Inter-Assay CV: <10% Kit Components: *355896A: Microtiter Strips, 1x48 wells or 1x96 wells *355896B: Standard, 1x1vial or 2x1 vial 355896C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml 355896D: Antibody (Biotin) (Concentrated), 1x60ul or 1x120ul 355896E: Antibody Dilution Buffer, 1x5ml or 1x10ml 355896F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul 355896G: SABC Dilution Buffer, 1x5ml or 1x10ml 355896H: TMB Substrate, 1x5ml or 1x10ml 355896J: Stop Solution, 1x5ml or 1x10ml 355896K: Wash Buffer, 25X, 1x15ml or 1x30ml Storage and Stability: Store unopened *355896A at 4C, store at -20C once opened. Store unopened *355896B 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 Principle: This ELISA kit employs the sandwich enzyme-linked immunoassay technique, utilizing a microtiter plate pre-coated with an antibody specific to GDF1. Standards and samples are added to the appropriate wells, then incubated. Aspirate and wash the plate. The Antibody (Biotin) is added to all the wells. The plate is incubated and then washed. Streptavidin (HRP) is added to each microplate well and incubated then washed. Then the TMB substrate solution is added and incubated. After the TMB substrate solution is added, only those wells that contain GDF1, the Antibody (Biotin) and Streptavidin (HRP) 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 GDF1 in the sample is then determined by comparing the O.D. of the sample to the standard curve. Assay Summary: 1. Wash plate 2 times before adding standards and samples to wells! 2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37C. Aspirate and wash 2 times. 3. Add 100ul Antibody (Biotin) working solution to each well and incubate for 60 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.
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