Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate ADAM17 in samples. An antibody specific for ADAM17 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyADAM17 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for ADAM17 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 ADAM17 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:ADAM17 is a 70-kDa enzyme that belongs to the ADAM protein family of disintegrins and metalloproteases. ADAM17 is understood to be involved in the processing of tumor necrosis factor alpha (TNF-alpha;) at the surface of the cell, and from within the intracellular membranes of the trans-Golgi network. This process, which is also known as 'shedding', involves the cleavage and release of a soluble ectodomain from membrane-bound pro-proteins (such as pro-TNF-alpha;), and is of known physiological importance.
ADAM17 was the first 'sheddase' to be identified, and is also understood to play a role in the release of a diverse variety of membrane-anchored cytokines, cell adhesion molecules, receptors, ligands, and enzymes.