特 异 性： 人MMP-3
检测标本： 细胞培养液、体液、血清和组织匀浆等
检测范围： 156pg/ml→10ng/ml
预期应用
ELISA法定量测定人血清、血浆、细胞培养物上清或其它相关液体中MMP-3含量。
实验原理
本试剂盒应用双抗体夹心酶标免疫分析法测定标本中MMP-3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-3抗原、生物素化的抗人MMP-3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-3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 MMP-3 concentrations in cell culture supernates, serum, and plasma.
Introduction
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, macroglobulins and tissue inhibitors of metalloproteinases (TIMPs).
MMP-3 is secreted from the cells as a proenzyme. The proenzyme has been shown to stimulate plasminogen activation. The N-terminal pro-domain contains the cysteine switch motif conserved in MMPs that maintains MMP-3 in the latent state. Activation of the proenzyme results in the removal of the pro-domain. MMP-3 activation can be achieved in vitro by proteases such as itself, chyrotrypsin, neutrophil elastase and plasma kallikrein, and by mercury compounds. The resulting active enzyme consists of a catalytic domain with a zinc-binding motif conserved in metzincins. A short hinge peptide links the catalytic domain to the C-terminal hemopexin-like domain. The active MMP-3 is capable of cleaving types III, IV, IX and X collagen, aggrecan, fibronectin, laminin, IGFBP-3, serpins, and IL-1 . The active enzyme also activates proMMP-1, -8, -9, and -13. Therefore, it is suggested that MMP-3 may participate in physiological matrix turnover and pathological destruction of the tissue. For example, MMP-3 is required for the generation of a macrophage chemoattractant in a model of herniated disc resorption.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for MMP-3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MMP-3 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for MMP-3 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-3 bound in the initial step. The color development is stopped and the intensity of the color is measured. |
