ELISA法定量测定小鼠血清、血浆或其它相关液体中MMP-13含量。

 

本试剂盒应用双抗体夹心酶标免疫分析法测定标本中MMP-13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MMP-13、生物素化的抗小鼠MMP-13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MMP-13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。

This immunoassay kit allows for the specific measurement of Mouse matrix metalloproteinase 13（MMP-13） concentrations in cell culture supernates, serum and plasma.

 

Matrix metalloproteinases (MMPs), also called matrixins, constitute a family of zinc and calcium

dependent endopeptidases that function in the breakdown of extracellular matrix (ECM). They play an important role in many normal physiological processes such as embryonic development, morphogenesis, reproduction and tissue remodeling . They also participate in many pathological processes such as arthritis, cancer and cardiovascular disease. While the amounts of newly synthesized MMPs are regulated mainly at the levels of transcription, the proteolytic activities of existing MMPs are controlled through both the activation of proenzymes or zymogens and the inhibition of active enzymes by endogenous inhibitors,-macroglobulin 2 and tissue inhibitors of metalloproteinases (TIMPs).

MMP-13 (collagenase 3) has been proposed to participate in aggrecan degradation associated with osteoarthritis and cleavage of type II collagen in osteoarthritic cartilage explants and in tumor progression and metastasis . In addition, it can cleave type I, III, IV, IX, X and XIV collagens and fibronectin . Therefore, MMP-13 is likely to play a crucial role in the modulation of extracellular matrix degradation and cell-matrix interactions .

MMP-13 is expressed by several metastatic tumors including breast carcinomas, chondrosarcomas, head and neck carcinomas, and in degenerative bone diseases including rheumatoid arthritis . Wild type and mutant p53, a tumor suppressor gene, differentially regulate MMP-13 gene expression . Members of the activator protein-1 and core-binding factor families increase MMP-13 promoter activity in normal, differentiating osteoblasts .

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for MMP-13 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MMP-13 present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for MMP-13 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 MMP-13 bound in the initial step. The color development is stopped and the intensity of the color is measured.