Each lot of this antibody is quality control tested by ELISA analysis. For ELISA capture applications, a concentration range of 5-10 μg/ml is recommended. To obtain a linear standard curve, serial dilutions of MCP-1 recombinant protein ranging from 2000 to 15 pg/ml are recommended for each ELISA plate. For immunofluorescent staining, the suggested use of this reagent is ≤0.5 μg per 106 cells in 100 μl volume. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
ELISA or ELISPOT Capture1: The purified 2H5 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated 4E2/MCP antibody (Cat. No. 506002) as the detecting antibody for the detection of mouse MCP-1. The purified 2H5 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated 5D3-F7 antibody (Cat. No. 502609) as the detecting antibody for the detection of human MCP-1. The LEAF™ purified antibody is suggested for ELISPOT capture. Neutralization1,4,6: The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for neutralization of mouse/rat MCP-1 bioactivity in vivo and in vitro (Cat. No. 505906). Additional reported applications (for the relevant formats) include: Western blotting3, immunohistochemistry5 of paraformaldehyde-fixed, saponin-treated frozen tissue sections.
Application References:
1. Luo, Y., et al., 1994. J. Immunol. 153: 3708. 2. Zhang, Y., et al., 2002. J. Immunol. 168: 3088. 3. Luo, Y., et al. 1999 J. Immunol. 163:3985. 4. Morrison, B. E., et al. 2003 J. Clin. Invest. 112:1862. 5. Hancock, W. W., et al. 1997 Transplantation 64:696. 6. Yu, R., et al. 2006 Obesity 14:1353. 7. Patsouris, D., et al. 2009. J. Biol Chem. 284:31223. PubMed.
Description:
Monocyte chemotactic protein-1 (MCP-1) also known as monocyte chemotactic and activating factor (MCAF) was identified based on its ability to chemoattract monocytes. Subsequently, MCP-1 has also been found to regulate adhesion molecule expression and cytokine production in monocytes. MCP-1 is identical to the product of the JE gene, a PDGF inducible gene. MCP-1 is a member of the beta (C-C) chemokine subfamily, known as CCL2. The 2H5 antibody reacts with mouse, rat, and human MCP-1. The 2H5 antibody can neutralize the bioactivity of natural or recombinant MCP-1.
Other Names:
Monocyte chemoattractant protein-1, Monocyte chemoattractant and activating factor (MCAF), JE, Small inducible cytokine A2 (SCYA2), HC-11, P6, Smooth muscle cell chemotactic factor (SMC-CF), CCL2,
Monocytes, basophils, T cells, NK cells, immature dendritic cells
Receptors:
CCR2; CCR5; CCR10; D6; possibly L-CCR
Bioactivity/Activities:
Chemoattractant; regulates adhesion molecule expression, cytokine production in monocytes; proliferation/activation of CC-chemokine activated killer cells
Antigen References:
1. Fitzgerald, K., et al., Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego. 2. Bischoff, S., et al., 1992. J. Exp. Med. 175:1271. 3. Charo, I., et al., 1994. PNAS 91:2752. 4. Jiang, Y., et al., 1992. J. Immunol. 148:2423.
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