Description
Principle of the Assay: The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the IgG2C present in samples reacts with the anti-IgG2C antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-IgG2C antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound IgG2C. Following another washing step, the enzyme bound to the immunosorbent 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 IgG2C in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of IgG2C in the test sample. The quantity of IgG2C in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution