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 microtitre wells. After the removal of unbound proteins by washing, anti-KIM-1 antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound KIM-1. 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 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.
KIM-1 is a trans-membrane structural glycoprotein in renal epithelial cells. These cells will shed KIM-1 antigen during regeneration into the urine which makes urinary KIM-1 a useful biomarker for kidney injury.

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