62.5-4000pg/ml

<37.5pg/ml

Intra-Assay CV: <8% Inter-Assay CV: <10%

This ELISA kit employs the sandwich enzyme-linked immunoassay technique, utilizing a microtiter plate pre-coated with an antibody specific to PINP. 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 PINP, 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 PINP in the sample is then determined by comparing the O.D. of the sample to the standard curve.

*358132A: Microtiter Strips, 1x48 wells or 1x96 wells *358132B: Standard, 1x1vial or 2x1 vial 358132C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml 358132D: Antibody (Biotin) (Concentrated), 1x60ul or 1x120ul 358132E: Antibody Dilution Buffer, 1x5ml or 1x10ml 358132F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul 358132G: SABC Dilution Buffer, 1x5ml or 1x10ml 358132H: TMB Substrate, 1x5ml or 1x10ml 358132J: Stop Solution, 1x5ml or 1x10ml 358132K: Wash Buffer, 25X, 1x15ml or 1x30ml

Store unopened *358132A at 4ºC; store at -20ºC once opened. Store unopened *358132B 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.

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 37°C. Aspirate and wash 2 times. 3. Add 100ul Antibody (Biotin) working solution to each well and incubate for 60 minutes at 37°C. 4. Aspirate and wash 3 times. 5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C 6. Aspirate and wash 5 times. 7. Add 90ul TMB substrate. Incubate 15-30 minutes at 37°C 8. Add 50ul Stop Solution. Read at 450nm immediately. 9. Calculate results