Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified 5D3-F7 antibody is useful as a capture antibody in a sandwich ELISA for measuring human MCP-1 protein levels. For specific methodology, please visit the protocols section of "ELISA and ELISPOT" on our web site, http://www.bdbiosciences.com/us/s/resources.
Note: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assay of serum or plasma samples. For measuring human MCP-1 in serum or plasma our Human MCP-1 BD OptEIA™ ELISA Set (Cat. No. 555179) or BD OptEIA ELISA Kit (Cat. No. 559107) are specially formulated and recommended.
Neutralization: The NA/LE™ 5D3-F7 antibody (Cat. No. 554661) has been reported to be useful for neutralization of human MCP-1 bioactivity.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 5D3-F7 monoclonal antibody specifically binds to human monocyte chemoattractant protein-1 (MCP-1), also known as CCL2 (C-C motif chemokine 2), Monocyte chemotactic and activating factor (MCAF), and Small-inducible cytokine A2 (SCYA2). MCP-1 is a 10-14 kDa glycoprotein member of the beta or CC family of chemokines. It expressed by monocytes, fibroblasts, endothelial cells and other cell types in response to IL-1, IL-6, TNF, and a variety of other stimuli. MCP-1 binds to and exerts its biological activity through G-protein coupled chemokine receptors including CCR2/CD192 and CCR4/CD194. It serves as a chemoattractant and activating factor for monocytes and other cell types including T cells, NK cells, and basophils.
MCP-1 is a member of the CC chemokine family and it is produced by monocytes, T lymphocytes, fibroblasts, endothelial cells, smooth muscle cells, keratinocytes and some tumors. Its production can be induced by LPS or IFN-γ. Clone 5D3-F7 also cross reacts with an intracellular component of LPS-stimulated (24 hours) peripheral blood monocytes of rhesus and cynomolgus macaque monkeys. The staining pattern observed on non-human primate monocytes is not as strong as that seen on normal human peripheral blood monocytes.
Development References (4)
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Peri G, Milanese C, Matteucci C, et al. 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 Methods. 1994; 174(1-2):249-257. (Clone-specific: Neutralization). View Reference
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Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: IC/FCM Block). View Reference
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Rollins BJ, Stier P, Ernst T, Wong GG. The human homolog of the JE gene encodes a monocyte secretory protein. Mol Cell Biol. 1989; 9(11):4687-4695. (Biology). View Reference
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Yoshimura T, Yuhki N, Moore SK, Appella E, Lerman MI, Leonard EJ. Human monocyte chemoattractant protein-1 (MCP-1). Full-length cDNA cloning, expression in mitogen-stimulated blood mononuclear leukocytes, and sequence similarity to mouse competence gene JE. FEBS Lett. 1989; 244(2):487-493. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
