Description
Principle of the Assay: alpha1beta GP ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-alpha1beta GP antibody and an alpha1beta GP-HRP conjugate. The assay sample and buffer are incubated together with alpha1beta GP-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the alpha1beta GP concentration since alpha1beta GP from samples and alpha1beta GP-HRP conjugate compete for the anti-alpha1beta GP antibody binding site. Since the number of sites is limited, as more sites are occupied by alpha1beta GP from the sample, fewer sites are left to bind alpha1beta GP-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The alpha1beta GP concentration in each sample is interpolated from this standard curve