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