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