*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.
Description
The 5D3-F7 antibody reacts with human, cynomolgus, and rhesus monkey monocyte chemoattractant protein-1 (MCP-1), also known as monocyte chemotactic and activating factor (MCAF) and CCL2. MCP-1 is a CC chemokine which presents as a dimer. Derived from monocytes and vascular endothelial cells, it exerts chemoattractant activity on a variety of cell types, such as monocytes, basophils, T cells and NK cells via chemokine receptors such as CCR2, CCR5, and CCR10.
Applications Reported
For research use only, not for diagnostic or therapeutic use. The 5D3-F7 antibody has been reported to be useful for detection of MCP-1 in a sandwich ELISA, neutralization of MCP-1 bioactivity, intracellular staining for flow cytometric analysis, IP, and immunohistochemical staining of frozen tissue sections.
Applications Tested
The affinity purified 5D3-F7 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of human MCP-1 in combination with the biotinylated 2H5 (13-7096) antibody for detection and recombinant human MCP-1 as the standard. A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 2000 pg/ml - 15 pg/ml should be included in each ELISA plate. The Functional Grade purified 5D3-F7 antibody has been tested by LAL assay to verify low endotoxin levels.
References
Peri, G., C. Milanese, C. Matteucci, C., L. Ruco, D. Zhou, S. Sozzani, I. Coletta, and A. Mantovani. 1994. A new monoclonal antibody (5D3-F7) which recognizes human monocyte-chemotactic protein-1 but not related chemokines. Development of a sandwich ELISA and in situ detection of producing cells. J. Immunol. Meth. 174: 249-257. (5D3-F7, ELISA, WB, IHC frozen),
Karrer S, Bosserhoff AK, et al. 2005. The -2518 promotor polymorphism in the MCP-1 gene is associated with systemic sclerosis. J Invest Dermatol. 124(1):92-8. (5D3-F7, IHC paraffin, PubMed)
Piemonti L, Leone BE, et al. 2002. Human pancreatic islets produce and secrete MCP-1/CCL2: relevance in human islet transplantation. Diabetes. 51(1):55-65. (5D3-F7, IHC paraffin, PubMed)
