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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MCP-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MCP-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MCP-1抗原、生物素化的抗大鼠MCP-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MCP-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 MCP-1 concentrations in cell culture supernates, serum, and plasma.
Introduction
Monocyte chemotactic protein 1 (MCP-1/CCL2), also known as monocyte chemotactic and activating factor (MCAF), is a member of the CC subgroup of the chemokine superfamily. As the name implies, MCP-1 is known for its ability to act as a potent chemoattractant and activator of monocytes/macrophages. Other cell types responding to MCP-1 include NK cells, T cells, eosinophils, basophils, and hepatic stellate cells. The human MCP-1 cDNA encodes a 99 amino acid (aa) residue precursor protein with a 23 aa hydrophobic signal peptide that is cleaved to generate the 76 aa mature protein. When compared to murine chemokines, MCP-1 is most closely related to MCP-5 (CCL12) and MCP-1/JE (CCL2). Murine MCP-5 shares 65% overall aa similarity with human MCP-1, while murine MCP-1/JE exhibits 62% aa identity excluding an unrelated 49 aa C-terminal sequence not found in the human protein. MCP-1 is induced by any inflammatory mediators such as LPS, IL-1 , TNF- , IFN- , IL-6, and IL-4. Several cell types have been shown to produce MCP-1 including mononuclear cells, mast cells, T cells, osteoblasts, fibroblasts, endothelial cells, bone marrow stromal cells, epithelial cells, microglia, astrocytes, and smooth muscle cells.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for MCP-1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MCP-1 present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for MCP-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 MCP-1 bound in the initial step. The color
development is stopped and the intensity of the color is measured. |
