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预期应用
ELISA法定量测定大鼠血清、血浆、细胞培养物上清或其它相关液体中MMP-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MMP-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MMP-1抗原、生物素化的抗大鼠MMP-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MMP-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 MMP-1 concentrations in cell culture supernates, serum, and plasma.
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- Introduction
- MMP-1, also known as collagenase, interstitial collagenase, vertebrate collagenase, and fibroblast collagenase, is produced by fibroblasts, chondrocytes, macrophages, endothelial cells, and osteoblasts. It is induced by the pro-inflammatory cytokines IL-1 and TNF-α, various growth factors such as EGF, PDGF, FGF basic, and Oncostatin M, chemical agents such as cAMP and phorbol esters and events occurring at the cell surface such as cell fusion and phagocytosis. MMP-1 plays a significant role in the degradation of different types of collagen in extracellular matrix remodeling, characterized by the cleavage of the interstitial collagen triple helix into ¾ and ¼ fragments. It is implicated in a variety of processes involving collagen degradation such as emphysema, atherosclerosis, rheumatoid and osteoarthritis, periodontal and respiratory disease, angiogenesis and tumorigenesis, tissue remodeling and wound healing, and inflammatory bowel disease. MMP-1 is capable of cleaving additional substrates such as casein, gelatin, aggrecan, entactin, pro-TNF, and cartilage link protein. Thus, the role of MMP-1 may be more diverse than originally envisaged.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for MMP-1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MMP-1 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for MMP-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 MMP-1 bound in the initial step. The color development is stopped and the intensity of the color is measured. |
