Description
For samples: Serum, plasma, cell culture supernatants, body fluid and tissue homogenate
INTENDED USE This C8b ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Human C8b. This ELISA kit for research use only, not for therapeutic or diagnostic applications!
PRINCIPLE OF THE ASSAY C8b ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-C8b antibody and an C8b-HRP conjugate. The assay sample and buffer are incubated together with C8b-HRP conjugat e inpre-coated plate for one hour. After the incubation period, the wells a redecanted 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 C8b concentration since C8b from samples and C8b-HRP conjugate compete for the anti-C8b antibody binding site. Since the number of sites is limited, as more sites are occupied by C8b from the sample, fewer sites are left to bind C8b-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The C8b concentration in each sample is interpolated from this standard curve