Rat Alpha 2 Macroglobulin ELISA

In this assay the alpha 2-macroglobulin (A2M) present in samples reacts with the anti-A2M antibodies which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, anti-A2M antibodies conjugated with horseradish peroxidase (HRP) are added. These enzyme-labeled antibodies form complexes with the previously bound A2M. Following another washing step, the bound enzyme 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 A2M in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of alpha 2-macroglobulin in the test sample. The quantity of A2M in the test sample can be interpolated from the standard curve constructed from the standards and corrected for sample dilution.