Tumor Necrosis Factor-alpha (TNF-alpha) BioAssay™ ELISA Kit (Hamster)

The Tumor Necrosis Factor-alpha (TNF-alpha) BioAssay™ ELISA Kit is a quantitative sandwich assay for the detection of TNF-alpha in hamster serum, plasma, tissue homogenates and other biological fluids.||Detection Range:|3.125-200pg/ml||Sensitivity:|1.875pg/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 TNF-alpha. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to TNF-alpha. 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 TNF-alpha, 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 TNF-alpha in the sample is then determined by comparing the O.D. of the sample to the standard curve.||Kit Components:|*370268A: Microtiter Strips, 1x48 wells or 1x96 wells |*370268B: Standard, 1x1vial or 2x1 vial|370268C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml|370268D: Antibody (Biotin) (Concentrated), 1x60ul or 1x120ul|370268E: Antibody Dilution Buffer, 1x5ml or 1x10ml|370268F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul|370268G: SABC Dilution Buffer, 1x5ml or 1x10ml|370268H: TMB Substrate, 1x5ml or 1x10ml|370268J: Stop Solution, 1x5ml or 1x10ml|370268K: Wash Buffer, 25X, 1x15ml or 1x30ml||Storage and Stability:|Store unopened *370268A at 4ºC; store at -20ºC once opened. Store unopened *370268B 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.||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 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 10-20 minutes at 37°C|8. Add 50ul Stop Solution. Read at 450nm immediately.|9. Calculate results