The 8-Isoprostane (8-iso-PGF2a) BioAssay™ ELISA Kit is a quantitative competitive assay for the detection of 8-Isoprostane in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples.||Detection Range:|15.625-1000pg/ml||Sensitivity:|9.375pg/ml||Precision: |Intra-Assay CV: <8%|Inter-Assay CV: <10%||Assay Principle:|This ELISA kit employs the quantitative competitive enzyme-linked immunoassay technique, utilizing a microtiter plate provided in this kit has been pre-coated with 8-iso-PGF2a. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to 8-iso-PGF2a. 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 8-iso-PGF2a, Biotin-conjugated antibody and enzyme-conjugated Streptavidin 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 8-iso-PGF2a in the sample is then determined by comparing the O.D. of the sample to the standard curve.||Kit Components:|*369693: Microtiter Strips, 1x48 wells or 1x96 wells |*369693B: Standard, 1x1 vial or 2x1 vials|369693C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml|369693D: Antibody (Biotin) (Concentrated), 1x1vial|369693E: Antibody Dilution Buffer, 1x5ml or 1x10ml|369693F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul|369693G: SABC Dilution Buffer, 1x5ml or 1x10ml|369693H: TMB Substrate, 1x5ml or 1x10ml|369693J: Stop Solution, 1x5ml or 1x10ml|369693K: Wash Buffer, 25X, 1x15ml or 1x30ml|369693L: Purified Water, 1x200ul||Storage and Stability:|Store unopened *369693A at 4ºC; store at -20ºC once opened. Store unopened *369693B 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 50ul standard or sample to each well.|3. Immediately add 50ul Antibody-Biotin working solution to each well and incubate for 45 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.