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