*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 Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
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 for neutralization of MCP-1 bioactivity.
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)
