Description
Intended Uses: This 5-HEPE ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Mouse 5-HEPE. This ELISA kit for research use only, not for therapeutic or test applications!
Principle of the Assay: 5-HEPE ELISA kit applies the competitive enzyme immunoassay technique utilizing an anti-5-HEPE antibody and an 5-HEPE-HRP conjugate. The assay sample and buffer are incubated together with 5-HEPE-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 5-HEPE concentration since 5-HEPE from samples and 5-HEPE-HRP conjugate compete for the anti-5-HEPE antibody binding site. Since the number of sites is limited, as more sites are occupied by 5-HEPE from the sample, fewer sites are left to bind 5-HEPE-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The 5-HEPE concentration in each sample is interpolated from this standard curve