The 8-Isoprostane （8-iso-PGF2a） BioAssay ELISA Kit （Rat） is a quantitative competitive assay for the detection of 8-Isoprostane in rat 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：369695A： Microtiter Strips，8x12 wells.369695B： Standard，2 vials369695C： Sample/Standard Dilution Buffer，1x20ml369695D： Antibody-Biotin Concentrate，1x160ul369695E： Antibody Dilution Buffer，1x10ml369695F： Streptavidin-HRP Conjugate （SABC），1x120ul369695G： SABC Dilution Buffer，1x10ml369695H： TMB Substrate，1x10ml369695J： Stop Solution，1x10ml369695K： Wash Buffer （25X），1x30mlStorage and Stability：Store unopened 369695A at 4°C; store at -20°C once opened. Store unopened 369695B at 4°C; once reconstituted，store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store 369695F and 369695H 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.
