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EK000057-EK0441: Human MCP-1 ELISA Kit
Range 15.6pg/ml-1000pg/ml
Sensitivity < 1 pg/ml
Specificity: cross-reactivates with human Eotaxin, MCP-3 <1%.
Application: for quantitive detection of human MCP-1 in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Monocyte chemoattractant protein-1 (MCP-1), a member of the chemokine (chemotactic cytokine) family, is a potent monocyte agonist that is upregulated by oxidized lipids.1 MCP-1 is also known as CCL2, SCYA2, MCAF. MCAF is a member of family of factors involved in immune and inflammatory responses. The amino acid sequence deduced from the nucleotide sequence reveals the primary structure of the MCAF precursor to be composed of a putative signal peptide sequence of 23 amino acid residues and a mature MCAF sequence of 76 amino acid residues.2 MCP-1 plays a unique and crucial role in the initiation of atherosclerosis and may provide a new therapeutic target in this disorder.3 Human MCP-1 is a 8.7KDa non-glycoprotein, consisting of 99 amino acids in precursor form and 76 amino acids in mature form.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Gosling, J.; Slaymaker, S.; Gu, L.; Tseng, S.; Zlot, C. H.; Young, S. G.; Rollins, B. J.; Charo, I. F. MCP-1 deficiency reduces susceptibility to atherosclerosis in mice that overexpress human apolipoprotein B. J. Clin. Invest. 103: 773-778, 1999.
2. Furutani, Y.; Nomura, H.; Notake, M.; Oyamada, Y.; Fukui, T.; Yamada, M.; Larsen, C. G.; Oppenheim, J. J.; Matsushima, K. Cloning and sequencing of the cDNA for human monocyte chemotactic and activating factor (MCAF). Biochem. Biophys. Res. Commun. 159: 249-255, 1989.
3. Gu, L.; Okaka, Y.; Clinton, S. K.; Gerard, C.; Sukhova, G. K.; Libby, P.; Rollins, B. J. Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. Molec. Cell 2: 275-281, 1998.
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