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The Human SPINT2 (HAI-2) ELISA quantitates Hu SPINT2 in human serum, plasma, or cell culture medium. The assay will exclusively recognize both natural and recombinant Hu SPINT2. Principle of the method The Human SPINT2 solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.HAI-2 is an endogenous inhibitor of the kunitz-type serine proteinase HGF-activator (HGFA). HGF was first described as a hepatocyte-specific mitogen and survival factor, and has since been shown to exert a variety of actions on many cell types by binding to its MET receptor. HGF is activated by cleavage of the single-chain form to form a two-chain version by HGFA. HAI-2 also inhibits trypsin, and MTSP-1 (Matriptase-1), a serine proteinase that also activates HGF, and HGF-2 has been shown to inhibit hepsin with nanamolar kIs. HAI-2 was first known as bikunin, and SPINT2 (Serine Proteinase Inhibitor-2), a proteinase inhibitor found in amniotic fluid. A protein termed HAI-2 related small protein is a 106 amino acid protein with little homology to HAI-2, and the HAI-2 antibodies do not recognize the HAI-2 RSP.|The chondroitin sulfate groups attached to HAI-2 are thought to be important for its function. The HAI-2 antibodies do not recognize HAI-1. Mice treated with HAI-2 in an ovarian cancer model had smaller tumors, and HAI-2 is reported to be elevated in macrophages during ovarian cancer, and this is considered a favorable indicator of survival. HAI-2 is secreted and membrane-inserted via a type-I transmembrane domain. The carboxyterminal end is a cytoplasmic domain, thought to interact with signaling and cytoskeletal machinery. The extracellular domains include two kringle domains (KD1 and KD2), and KD2 is thought to be inactive or with much lower activity against HGF. In mice the dominant HAI-2 lacks the first kringle domain, but the two-domain form dominates in humans, although in human testis a shorter HAI-2 has been reported, using a second transcription site.