Description
Intended Uses: This SEMA4D ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Sheep SEMA4D. This ELISA kit for research use only, not for therapeutic or diagnostic applications!
Principle of the Assay: SEMA4D ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-SEMA4D antibody and an SEMA4D -HRP conjugate. The assay sample and buffer are incubated together with SEMA4D -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 SEMA4D concentration since SEMA4D from samples and SEMA4D -HRP conjugate compete for the anti-SEMA4D antibody binding site. Since the number of sites is limited, as more sites are occupied by SEMA4D from the sample, fewer sites are left to bind SEMA4D -HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The SEMA4D concentration in each sample is interpolated from this standard curve