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/ml||Sensitivity:|<19.375pg/ml||Precision: |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 vials|369695C: Sample/Standard Dilution Buffer, 1x20ml|369695D: Antibody-Biotin Concentrate, 1x160ul|369695E: Antibody Dilution Buffer, 1x10ml|369695F: Streptavidin-HRP Conjugate (SABC), 1x120ul|369695G: SABC Dilution Buffer, 1x10ml|369695H: TMB Substrate, 1x10ml|369695J: Stop Solution, 1x10ml|369695K: Wash Buffer (25X), 1x30ml||Storage 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°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 15-20 minutes at 37°C|8. Add 50ul Stop Solution. Read at 450nm immediately.|9. Calculate results.