Rat IgG ELISA

In this assay the IgG present in samples reacts with the anti-IgG antibodies which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, antiIgG antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound IgG. Following another washing step, the bound enzyme-labeled anti-IgG antibody is assayed by the addition of a chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of IgG in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of IgG in the test sample. The quantity of IgG in the test sample can be interpolated from the standard curve constructed from the standards and corrected for sample dilution.