Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate A2ML1 in samples. An antibody specific for A2ML1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyA2ML1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for A2ML1 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 A2ML1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:The alpha-macroglobulin (AM) superfamily of proteins contains both complement components and protease inhibitors, including A2M and A2ML1. AM proteins display a unique trap mechanism of inhibition, by which the AM inhibitor undergoes a major conformational change upon its cleavage by a protease, thus trapping the protease and blocking it from subsequent substrate binding. This protein has a peptide stretch, called the 'bait region' which contains specific cleavage sites for different proteinases. When a proteinase cleaves the bait region, a conformational change is induced in the protein which traps the proteinase. The entrapped enzyme remains active against low molecular weight substrates. Following cleavage in the bait region a thioester bond is hydrolyzed and mediates the covalent binding of the protein to the proteinase.
