Free Haemoglobin (f-Hb) BioAssay(TM) ELISA Kit (Human)

Artikelnummer: USB-355523
Artikelname: Free Haemoglobin (f-Hb) BioAssay(TM) ELISA Kit (Human)
Artikelnummer: USB-355523
Hersteller Artikelnummer: 355523
Alternativnummer: USB-355523-48, USB-355523-96
Hersteller: US Biological
Kategorie: Kits/Assays
The Free Haemoglobin (f-Hb) BioAssay(TM) ELISA Kit is a quantitative sandwich assay for the detection of f-Hb in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples. Detection Range: 0.313-20ng/ml Sensitivity: 0.188ng/ml Precision: Intra-Assay: CV<8% Inter-Assay: CV<10% Assay Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to f-Hb. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to f-Hb. 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 f-Hb, Biotin-conjugated antibody and enzyme-conjugated Streptavidin complex 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 f-Hb in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *355523A: Microtiter Strips, 1x48 wells or 1x96 wells *355523B: Standard, 1x1 vial or 2x1 vial 355523C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml 355523D: Antibody (Biotin) (Concentrated), 1x60ul or 1x120ul 355523E: Antibody Dilution Buffer, 1x5ml or 1x10ml 355523F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul 355523G: SABC Dilution Buffer, 1x5ml or 1x10ml 355523H: TMB Substrate, 1x5ml or 1x10ml 355523J: Stop Solution, 1x5ml or 1x10ml 355523K: Wash Buffer, 25X, 1x15ml or 1x30ml Storage and Stability: Store unopened *355523A at 4C, store at -20C once opened. Store unopened *355523B 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 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