Description
Principle of the Assay: TCF 20 ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-TCF 20 antibody and an TCF 20-HRP conjugate. The assay sample and buffer are incubated together with TCF 20-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 TCF 20 concentration since TCF 20 from samples and TCF 20-HRP conjugate compete for the anti-TCF 20 antibody binding site. Since the number of sites is limited, as more sites are occupied by TCF 20 from the sample, fewer sites are left to bind TCF 20-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The TCF 20 concentration in each sample is interpolated from this standard curve