Description
Introduction: Tumor necrosis factor (TNF, cachexin or cachectin and formally known as tumor necrosis factor-alpha) is a cytokine involved in systemic inflammation and is a member of a group of cytokines that stimulate the acute phase reaction.The primary role of TNF is in the regulation of immune cells. TNF is also able to induce apoptotic cell death, to induce inflammation, and to inhibit tumorigenesis and viral replication. Dysregulation and, in particular, overproduction of TNF have been implicated in a variety of human diseases, as well as cancer.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to TNF-alpha. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for TNF-alpha and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain TNF-alpha, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of TNF-alpha in the samples is then determined by comparing the O.D. of the samples to the standard curve