预期应用
ELISA法定量测定大鼠血清、血浆或其它相关液体中基质金属蛋白酶抑制因子-1(TIMP-1)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TIMP-1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TIMP-1抗原、生物素化的抗大鼠TIMP-1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TIMP-1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 Rat tissue inhibitors of metalloproteinase,TIMP-1 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 and in the processing of a variety of biological molecules. 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 such as-macroglobulins and tissue inhibitors of α2 metalloproteinases (TIMPs).
The mammalian TIMP family includes four members that share structural similarity.All TIMP proteins have 12 conserved cysteine residues that form six intrachain disulfide bonds, resulting in an extremely stable protein with six loops. The TIMP protein has two structurally and functionally distinct domains: the N-terminal domain consisting of loops 1 - 3 that are responsible for tight, but non-covalent binding to the active MMPs in a 1:1 stoichiometry; and the C-terminal domain consisting of loops 4 - 6 that enhances the enzyme-inhibitor interactions. In the case of TIMP-1, the C-terminal domain has also been shown to bind the hemopexin-like domain of pro MMP-9. TIMP-1 stimulates erythropoiesis, inhibits angiogenesis and is an anti-apoptotic agent for B cells. These TIMP-1 functions may be independent of MMP inhibition .
Rat TIMP-1 is a 28 - 35 kDa secreted glycoprotein . The protein is synthesized as a 217 amino acid (aa) precursor that contains a 23 aa signal peptide and a 194 aa mature form. TIMP-1 is widely expressed in many cells such as fibroblasts, osteoblasts, endothelial cells, granulosa cells, dendritic cells, vascular smooth muscle cells, adipocytes, and monocytes.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for TIMP-1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TIMP-1 present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for TIMP-1 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 TIMP-1 bound in the initial step. The color development is stopped and the intensity of the color is measured. | 
