Description
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to Steroid 21-hydroxylase. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for Steroid 21-hydroxylase and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain Steroid 21-hydroxylase, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of Steroid 21-hydroxylase in the samples is then determined by comparing the O.D. of the samples to the standard curve