PIG3 (p53 inducible gene 3)

The PIG3 ELISA is a sandwich enzyme immunoassay employing mouse monoclonal antibodies. A monoclonal antibody, specific for the human PIG3 protein, has been immobilized onto the surface of microtiter wells provided in the kit. The sample to be assayed and the biotinylated detector monoclonal antibody are pipetted into the wells and allowed to incubate for two hours, during which time any PIG3 present binds to the capture and detecting antibodies. Unbound material is washed away and horseradish peroxidase-conjugated streptavidin is added, which binds to the detector antibody.The horseradish peroxidase catalyzes the conversion of the chromogenic substrate tetra-methylbenzidine (TMB) from a colorless solution to a blue solution (or yellow after the addition of stop reagent), the intensity of which is proportional to the amount of PIG3 protein in the sample. The colored reaction product is quantified using a spectrophotometer.Quantitation is achieved by the construction of a standard curve using known concentrationsof PIG3 (provided lyophilized). By comparing the absorbance obtained from a sample containing an unknown amount of PIG3 with that obtained from the standards, the concentration of PIG3 in the sample can be determined.