Description
Principle of the Assay: Caspase 14 ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-Caspase 14 antibody and an Caspase 14-HRP conjugate. The assay sample and buffer are incubated together with Caspase 14-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 Caspase 14 concentration since Caspase 14 from samples and Caspase 14-HRP conjugate compete for the anti-Caspase 14 antibody binding site. Since the number of sites is limited, as more sites are occupied by Caspase 14 from the sample, fewer sites are left to bind Caspase 14-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The Caspase 14 concentration in each sample is interpolated from this standard curve