Description
Principle of the assay: Ras protein ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-Ras protein antibody and an Ras protein-HRP conjugate. The assay sample and buffer are incubated together with Ras protein-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 Ras protein concentration since Ras protein from samples and Ras protein-HRP conjugate compete for the anti-Ras protein antibody binding site. Since the number of sites is limited, as more sites are occupied by Ras protein from the sample, fewer sites are left to bind Ras protein-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The Ras protein concentration in each sample is interpolated from this standard curve