H-2 Class II Histocompatibility Antigen, A-B alpha chain (H2-Aa) BioAssay(TM) ELISA Kit (Mouse)
Artikelnummer:
USB-356180
Hersteller Artikelnummer:
356180
Alternativnummer:
USB-356180-48, USB-356180-96
Hersteller:
US Biological
Kategorie:
Kits/Assays
The H-2 class II histocompatibility antigen, A-B alpha chain (H2-Aa) BioAssay(TM) ELISA Kit s a quantitative sandwich assay for the detection of H2-Aa in mouse serum, plasma, tissue homogenates and other biological fluids. Detection Range: 0.625-40ng/ml Sensitivity: 0.375ng/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 H2-Aa. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to H2-Aa. 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 H2-Aa, 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 H2-Aa in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *356180A: Microtiter Strips, 1x48 wells or 1x96 wells *356180B: Standard, 1x1 vial or 2x1 vial 356180C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml 356180D: Antibody-Biotin (Concentrated), 1x60ul or 1x120ul 356180E: Antibody Dilution Buffer, 1x5ml or 1x10ml 356180F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul 356180G: SABC Dilution Buffer, 1x5ml or 1x10ml 356180H: TMB Substrate, 1x5ml or 1x10ml 356180J: Stop Solution, 1x5ml or 1x10ml 356180K: Wash Buffer, 25X, 1x15ml or 1x30ml Storage and Stability: Store unopened *356180A at 4C, store at -20C once opened. Store unopened *356180B 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, samples and control (zero) 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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