Description
The principle of the following enzyme immunoassay test follows the typical competitive binding scenario. Competition occurs between an unlabelled estradiol (present in standards, controls and patient samples) and an enzyme-labelled estradiol (conjugate) for a limited number of antibody binding sites on the microplate. The washing and decanting procedures remove unbound materials. After the washing step, the enzyme substrate is added. The enzymatic reaction is terminated by addition of the stopping solution. The absorbance is measured on a microtiter plate reader. The intensity of the colour formed is inversely proportional to the concentration of unlabeled estradiol in the sample. A set of standards is used to plot a standard curve from which the amount of estradiol in patient samples and controls can be directly read