Description
Principle of the Assay: OVA sIgG1 ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-OVA sIgG1 antibody and an OVA sIgG1-HRP conjugate. The assay sample and buffer are incubated together with OVA sIgG1-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 OVA sIgG1 concentration since OVA sIgG1 from samples and OVA sIgG1-HRP conjugate compete for the anti-OVA sIgG1 antibody binding site. Since the number of sites is limited, as more sites are occupied by OVA sIgG1 from the sample, fewer sites are left to bind OVA sIgG1-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The OVA sIgG1 concentration in each sample is interpolated from this standard curve