Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified monoclonal antibody B8-11 is useful as a capture antibody in a sandwich ELISA for measuring human MIG protein levels. Purified monoclonal antibody B8-11 can be paired with the biotinylated monoclonal antibody B8-6 (Cat. No. 555037) with recombinant human MIG (Cat. No. 554636) as the standard. Purified monoclonal antibody B8-11 should be titrated to determine optimal concentration for ELISA capture (0.5-4.0 µg/ml). To obtain linear standard curves, doubling dilutions of human MIG ranging from ~2,500 to 39 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit the protocols section or chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.
This ELISA pair shows no cross-reactivity with any of the cytokines tested ( human IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-13, IL- 15, IL-16, eotaxin, G-CSF, GM-CSF, GROα, GROβ, GROγ, IFN-γ, IP-10, lymphotactin, MCP-1, MCP-2, MCP-3, MCP-4, MIP-1α, MIP-1β, NAP-2, PF-4, RANTES, TNF, LT-α).
This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assay of serum samples. The BD OptEIA™ ELISA set (Cat. No. 550998) is specially-formulated for cytokine measurement in complex fluids such as serum and plasma.
Immunofluorescent Staining and Flow Cytometric Analysis: The B8-11antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify and enumerate MIG producing cells within mixed cell populations. The PE-conjugated B8-11 antibody (Cat. No. 566013) is especially suitable for these studies.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
The B8-11 monoclonal antibody specifically recognizes monokine induced by gamma interferon (MIG), which is also known as chemokine (C-X-C motif) ligand 9 (CXCL9). MIG is a chemotactic cytokine that belongs to the CXC subfamily of chemokines. MIG is predominantly expressed by monocytes, macrophages, hepatocytes, and endothelial cells in response to interferon-gamma (IFN-γ) stimulation. It is primarily involved in the recruitment of activated T cells. MIG/CXCL9, and the closely-related IP10/CXCL10 and I-TAC/CXCL11 chemokines, exert their biological activities by binding to and signaling through the CXCR3 (CD183) chemokine receptor. CXCR3 is expressed on multiple cell types but predominantly on memory and effector T cells. The CXCL9 and CXCR3 interaction plays an important role in the onset of inflammation and is implicated in the pathogenesis of T-cell mediated immunity in several disease models.
Development References (4)
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Farber JM. A macrophage mRNA selectively induced by gamma-interferon encodes a member of the platelet factor 4 family of cytokines. Proc Natl Acad Sci U S A. 1990; 87(14):5238-5242. (Biology). View Reference
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Farber JM. HuMig: a new human member of the chemokine family of cytokines. Biochem Biophys Res Commun. 1993; 192(1):223-230. (Biology). View Reference
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Liao F, Rabin RL, Yannelli JR, Koniaris LG, Vanguri P, Farber JM. Human Mig chemokine: biochemical and functional characterization. J Exp Med. 1995; 182(5):1301-1314. (Biology). 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. (Clone-specific: Flow cytometry, IC/FCM Block, Immunofluorescence). View Reference
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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.