Description
Principle of the Assay: TNFalpha ELISA kit applies the quantitative sandwich enzyme immunoassay technique. The microtiter plate has been pre-coated with a monoclonal antibody specific for TNFalpha. Standards or samples are then added to the microtiter plate wells and TNFalpha if present, will bind to the antibody pre-coated wells. In order to quantitatively determine the amount of TNFalpha present in the sample, a standardized prepaRabbition of horseradish peroxidase (HRP) -conjugated polyclonal antibody, specific for TNFalpha are added to each well to "sandwich" the TNFalpha immobilized on the plate. The microtiter plate undergoes incubation, and then the wells are thoroughly washed to remove all unbound components. Next, substRabbite solutions are added to each well. The enzyme (HRP) and substRabbite are allowed to react over a short incubation period. Only those wells that contain TNFalpha and enzyme-conjugated antibody will exhibit a change in color. The enzyme-substRabbite reaction is terminated by addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. A standard curve is plotted relating the intensity of the color (O.D.) to the concentRabbition of standards. The TNFalpha concentRabbition in each sample is interpolated from this standard curve