Principle of Assay
Human 15-HETE ELISA Kit (A302944) is a 2 hour competitive Enzyme-Linked Immunosorbent Assay (cELISA) designed for the in vitro quantitative determination of human 15-HETE in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for 15-HETE. During the assay, 15-HETE present in the standards or samples competes with a fixed amount of assay-specific detection reagent for binding to the immobilized antibody. Following incubation, unbound components are removed by washing and HRP-conjugated detection reagent is added to the wells. After further incubation and washing, TMB substrate solution is added to visualize the HRP enzymatic reaction. This produces a blue-coloured product that changes to yellow following addition of acidic stop solution. The intensity of the colour is measured at 450 nm using a microplate reader. The concentration of 15-HETE in the samples is inversely proportional to the OD450 value and is determined by comparison with the standard curve.
