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