Canine KIM-1 ELISA

In this assay, the KIM-1 present in samples reacts with the anti-KIM-1 antibodies which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, the detection antibody, biotin conjugated anti-KIM-1, is added and complexes are formed. Following washing step, the horseradish peroxidase (HRP) conjugated streptavidin is added and complexes are formed. After another washing step, the complexes are 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 KIM-1 in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of KIM-1 in the test sample. The quantity of KIM-1 in the test sample can be interpolated from the standard curve constructed from the standards and corrected for sample dilution.