Description
This assay employs the competitive enzyme immunoassay technique. A specific anti-mouse-IgG antibody has been pre-coated onto a microtiter plate. Cortisol containing samples or standards and a Cortisol -HRP conjugate are given into the wells of the microtiter plate. Then a mouse monoclonal antibody to Cortisol is added into the wells. Cortisol in samples or standards and a fixed amount of Cortisol -HRP conjugate are competing for a limited amount of Cortisol mouse monoclonal antibody. The Cortisol-antibody complex binds tothe goat polyclonal anti-mouse IgG that has been previously bound to the well. After incubation, the wells are washed with wash buffer to remove unbound material. A substrate solution is then added and incubated, resulting in the development of a blue color. The color development is inhibited by the addition of a stop solution, and the color turns yellow. The yellow color is measured at 450 nm. The concentration of Cortisol is indirectly proportional to the color intensity of the test sample