Description
Intended Uses: This APAb ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Monkey APAb. This ELISA kit for research use only, not for therapeutic or test applications!
Principle of the Assay: APAb ELISA kit applies the competitive enzyme immunoassay technique utilizing phospholipid antigen and an APAb-HRP conjugate. The assay sample and buffer are incubated together with APAb-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 APAb concentration since APAb from samples and APAb-HRP conjugate compete for the phospholipid antigen binding site. Since the number of sites is limited, as more sites are occupied by APAb from the sample, fewer sites are left to bind APAb-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The APAb concentration in each sample is interpolated from this standard curve