For the direct quantitative determination of 3α Diol G by enzyme immunoassay in human serum. The principle of the following enzyme immunoassay test follows the typical competitive binding scenario. Competition occurs between an unlabeled antigen (present in standards, controls and patient samples) and an enzyme-labelled antigen (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 color formed is inversely proportional to the concentration of 3α Diol G in the sample. A set of standards is used to plot a standard curve from which the amount of 3α Diol G in patient samples and controls can be directly read.