Description
Principle of the Assay: The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Alpha 2-Macroglobulin present in samples reacts with the anti-Alpha 2-Macroglobulin antibodies which have been adsorbed to the surface of polystyrene microtitre 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 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 A2M in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of A2M 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.
Background: Alpha 2-Macroglobulin (A2M) is a major protease inhibitor in serum and an acute phase protein which increases significantly in concentration in the rat as a result of inflammation. The major pathophysiological role for rat alpha 2-macroglobulin has yet to be conclusively defined. This kit is specific to the alpha 2- macroglobulin and will not cross react with the closely related alpha 1-macroglobulin