Description
Principle of the assay: This assay employs a two-site sandwich ELISA to quantitative TNF-alpha in Rat serum, plasma, tissue homogenates. An antibody specific for TNF-alpha has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TNF-alpha present is bound by the immobilized antibody. After removing any unbound substances, a biotin - conjugated antibody specific for TNF-alpha is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TNF-alpha bound in the initial step. The color development is stopped and the intensity of the color is measured.
Background: Tumor necrosis factor (TNF, cachexin, or cachectin, and formerly known as tumor necrosis factor alpha or TNF-alpha) is an adipokine involved in systemic inflammation and is a member of a group of cytokines that stimulate the acute phase reaction. It is produced chiefly by activated macrophages, although it can be produced by many other cell types such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons.The primary role of TNF is in the regulation of immune cells. TNF, being an endogenous pyrogen, is able to induce fever, apoptotic cell death, cachexia, inflammation and to inhibit tumorigenesis and viral replication and respond to sepsis via IL1 & IL6 producing cells. Dysregulation of TNF production has been implicated in a variety of human diseases including Alzheimer is disease, cancer, major depression and inflammatory bowel disease (IBD). While still controversial, studies of depression and IBD are currently being linked to TNF levels. Recombinant TNF is used as an immunostimulant under the INN tasonermin. TNF can be produced ectopically in the setting of malignancy and parallels parathyroid hormone both in causing secondary hypercalcemia and in the cancers with which excessive production is associated