Rat C-Reactive Protein ELISA (Rat CRP ELISA)

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