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