Description
For samples: Serum, plasma, cell culture supernatants, body fluid and tissue homogenate
INTENDED USE This PCNA ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Rat PCNA. This ELISA kit for research use only, not for therapeutic or diagnostic applications!
PRINCIPLE OF THE ASSAY PCNA ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-PCNA antibody and an PCNA-HRP conjugate. The assay sample and buffer are incubated together with PCNA-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 PCNA concentration since PCNA from samples and PCNA-HRP conjugate compete for the anti-PCNA antibody binding site. Since the number of sites is limited, as more sites are occupied by PCNA from the sample, fewer sites are left to bind PCNA-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The PCNA concentration in each sample is interpolated from this standard curve