Description
Principle of the assay: PDHE1 ELISA kit applies the competitive enzyme immunoassay technique utilizing a PDHE1 and an PDHE1-HRP conjugate. The assay sample and buffer are incubated together with PDHE1-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 450 nm in a microplate reader. The intensity of the color is inversely proportional to the PDHE1 concentration since PDHE1 from samples and PDHE1-HRP compete for the PDHE1 antibody binding site. Since the number of sites is limited, as more sites are occupied by PDHE1 from the sample, fewer sites are left to bind PDHE1-HRP. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The PDHE1 concentration in each sample is interpolated from this standard curve