本试剂盒应用双抗体夹心酶标免疫分析法测定标本中ADAM8水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入ADAM8抗原、生物素化的抗人ADAM8抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的ADAM8呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。
Intended use
This immunoassay kit allows for the specific measurement of human ADAM8 concentrations in serum and plasma.
Introduction
The ADAM (A Disintegrin And Metalloprotease) gene family encodes a group of proteins with a common domain structure including a pro-, metalloprotease, disintegrin-like, cysteine-rich, transmembrane and cytoplasmic domain. The ADAMs function in many important processes such as fertilization and development through their activities in cell adhesion/fusion, membrane protein shedding, and possibly signal transduction. About half of the known ADAMs, including ADAM8, are predicted to contain a functional zinc-binding site and thus, said to be active metalloproteases. ADAM8, also known as cell surface antigen CD156 and MS2, is expressed in macrophages, granulocytes, monocytes and B cells. It is an osteoclast stimulating factor. ADAM8 may play a role in neuron-glia interactions during neurodegeneration. It may also mediate extravasation of leukocytes either by degrading the vascular basement membrane or by liberating active molecules from their precursors harbored in the cell surface of leukocytes and endothelial cells. Human ADAM8 precursor consists of 824 amino acid (aa) residues consisting of a 16 aa signal peptide, a 637 aa ectodomain including pro-, metalloprotease, disintegrin-like and cysteine-rich regions, a 25 aa transmembrane domain, and a 146 aa cytoplasmic domain). ADAM8 naturally occurs as both a membrane and a soluble form. It is an active protease, capable of cleaving myelin basic protein and a variety of peptide substrates based on the cleavage sites of membrane-bound cytokines, growth factors and receptors that are known to be processed by metalloproteases.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for ADAM8 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ADAM8 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for ADAM8 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of ADAM8 bound in the initial step. The color development is stopped and the intensity of the color is measured.
