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