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