Description
For samples: Serum, plasma, cell culture supernatants, body fluid and tissue homogenate
INTENDED USE This 4-HNE ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Sheep 4-HNE. This ELISA kit for research use only, not for therapeutic or diagnostic applications!
PRINCIPLE OF THE ASSAY 4-HNE ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-4-HNE antibody and an 4-HNE-HRP conjugate. The assay sample and buffer are incubated together with 4-HNE-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 4-HNE concentration since 4-HNE from samples and 4-HNE-HRP conjugate compete for the anti-4-HNE antibody binding site. Since the number of sites is limited, as more sites are occupied by 4-HNE from the sample, fewer sites are left to bind 4-HNE-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The 4-HNE concentration in each sample is interpolated from this standard curve