Description
Principle of the Assay: The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Alpha 1 Microglobulin present in samples reacts with the anti-Alpha 1 Microglobulin antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-A1M antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound A1M. 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 A1M in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of A1M in the test sample. The quantity of A1M in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.
Background: Alpha 1 Microglobulin (A1M) is a small molecular weight glycoprotein. It is heterogeneous in charge and can occur both as a monomer or a polymer. Alpha 1 Microglobulin is produced in the liver and can be used as indicator for some renal diseases