Description
Intended Uses: This HLA-C ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Human HLA-C. This ELISA kit for research use only!
Principle of the Assay: HLA-C ELISA kit applies the competitive enzyme immunoassay technique utilizing an anti-HLA-C antibody and an HLA-C-HRP conjugate. The assay sample and buffer are incubated together with HLA-C-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 HLA-C concentration since HLA-C from samples and HLA-C-HRP conjugate compete for the anti-HLA-C antibody binding site. Since the number of sites is limited, as more sites are occupied by HLA-C from the sample, fewer sites are left to bind HLA-C-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The HLA-C concentration in each sample is interpolated from this standard curve