The 4-Hydroxynonenal （4-HNE） BioAssay ELISA Kit is a quantitative competitive assay for the detection of 4-HNE in mouse serum，plasma，tissue homogenates and other biological fluids.Detection Range：31.2-2000pg/mlSensitivity：<18.75pg/mlPrecision：Intra-assay CV： <8%Inter-assay CV： <10%Assay Principle：The microtiter plate provided in this kit has been pre-coated with 4-HNE. 4-HNE in the standard or sample competes with a fixed amount of plate-coated 4-HNE for antibody binding sites on the Antibody-Biotin reagent. Excess conjugate and unbound sample or standard are washed from the plate，after which Streptavidin conjugated to Horseradish Peroxidase （HRP） is added to each microplate 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 4-HNE in the samples is determined by an inverse correlation between 4-HNE in the sample and the assay signal intensity.Kit Components：*369688A： Microtiter Strips，1x96 wells （8x12 wells）.*369688B： Standard，2x1 vial369688C： Sample/Standard Dilution Buffer，1x20ml369688D： Antibody-Biotin （Concentrated），1x60ul369688E： Antibody Dilution Buffer，1x10ml369688F： Streptavidin （HRP） （SABC，1x120ul369688G： SABC Dilution Buffer，1x10ml369688H： TMB Substrate，1x10ml369688J： Stop Solution，1x10ml369688K： Wash Buffer，25X，1x30mlStorage and Stability：Store unopened *369688A at 4ºC; store at -20ºC once opened. Store unopened *369688B 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°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.
