The 8-Isoprostane (8-iso-PGF2a) BioAssay ELISA Kit (Human) is a quantitative competitive assay for the detection of 8-Isoprostane in human serum,plasma,tissue homogenates and other biological fluids.Detection Range:15.25-1,000pg/mlSensitivity:<19.375pg/mlPrecision: Intra-Assay CV: <8%Inter-Assay CV: <10%Assay Principle:The microtiter plate provided in this kit has been pre-coated with 8-Isoprostane. 8-Isoprostane in the standard or sample competes with a fixed amount of plate-coated 8-Isoprostane for antibody binding sites on the Antibody-Biotin reagent. Excess conjugate and unbound sample or standard are washed from the plate,after which Streptavidin-HRP (SABC) is added to each well and incubated. TMB substrate is then added to each well. The enzyme-substrate reaction is terminated by the addition of a sulfuric acid solution and the resulting color is measured spectrophotometrically at 450nm. The concentration of 8-Isoprostane in the samples is determined by comparing the absorbance of the samples to the standard curve.Kit Components:369693A: Microtiter Strips,8x12 wells.369693B: Standard,2 vials369693C: Sample/Standard Dilution Buffer,1x20ml369693D: Antibody-Biotin Concentrate,1x160ul369693E: Antibody Dilution Buffer,1x10ml369693F: Streptavidin-HRP Conjugate (SABC),1x120ul369693G: SABC Dilution Buffer,1x10ml369693H: TMB Substrate,1x10ml369693J: Stop Solution,1x10ml369693K: Wash Buffer (25X),1x30mlStorage and Stability:Store unopened 369693A at 4°C; store at -20°C 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 369693F and 369693H protected from light at 4°C. Store the rest of the 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,samples and control (zero) 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°C4. Aspirate and wash 3 times.5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate and wash 5 times.7. Add 90ul TMB substrate. Incubate 15-20 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately.9. Calculate results.