Principle of Assay
Universal Estradiol ELISA Kit (A335449) is a 120 minute competitive Enzyme-Linked Immunosorbent Assay (cELISA) designed for the in vitro quantitative determination of universal Estradiol in serum, plasma, cell culture supernatant, cell or tissue lysate, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for Estradiol. During the assay, Estradiol 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 Estradiol in the samples is inversely proportional to the OD450 value and is determined by comparison with the standard curve.
