Flow cytometric analysis of MIG (CXCL9) expression in activated human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) were cultured overnight without (Unstimulated; Left Panel) or with (IFN-γ and TNF Stimulated; Right Panel) BD Pharmingen™ Recombinant Human IFN-γ (50 ng/mL; Cat. No. 554616) and Recombinant Human TNF (50 ng/mL; Cat. No. 554618) proteins. BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724) was added to the cell cultures four hours before fixing and permeabilizing the cells with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histograms) or PE Mouse Anti-Human MIG (CXCL9) antibody (Cat. No. 566013; solid line histograms). The overlaid histograms showing the expression of MIG (CXCL9) or Ig Isotype control staining were derived from gated events with forward and side light-scatter characteristics of intact monocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
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Flow cytometric analysis of MIG (CXCL9) expression in activated human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) were cultured overnight without (Unstimulated; Left Panel) or with (IFN-γ and TNF Stimulated; Right Panel) BD Pharmingen™ Recombinant Human IFN-γ (50 ng/mL; Cat. No. 554616) and Recombinant Human TNF (50 ng/mL; Cat. No. 554618) proteins. BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724) was added to the cell cultures four hours before fixing and permeabilizing the cells with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histograms) or PE Mouse Anti-Human MIG (CXCL9) antibody (Cat. No. 566013; solid line histograms). The overlaid histograms showing the expression of MIG (CXCL9) or Ig Isotype control staining were derived from gated events with forward and side light-scatter characteristics of intact monocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
C-X-C motif chemokine 9; CMK; crg-10; Humig; SCYB9
Human (QC Testing)
Mouse IgG1, κ
Recombinant Human MIG Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2739458
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation and Permeabilization Solution RUO
Size
125 mL
Cat No.
554722
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 mL
Cat No.
554724
PE Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554680
Recombinant Human IFN-γ RUO
Size
25 µg
Cat No.
554616
566013 Rev. 1
Antibody Details
B8-11
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.
566013 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566013 Rev.1
Citations & References
View product citations for antibody "566013" on CiteAb
Development References (9)
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Albanesi C, Scarponi C, Sebastiani S, et al. IL-4 enhances keratinocyte expression of CXCR3 agonistic chemokines.. J Immunol. 2000; 165(3):1395-402. (Clone-specific: ELISA). View Reference
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Bendriss-Vermare N, Burg S, Kanzler H, et al. Virus overrides the propensity of human CD40L-activated plasmacytoid dendritic cells to produce Th2 mediators through synergistic induction of IFN-{gamma} and Th1 chemokine production.. J Leukoc Biol. 2005; 78(4):954-66. (Clone-specific: Flow cytometry). View Reference
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Brice GT, Graber NL, Hoffman SL, Doolan DL. Expression of the chemokine MIG is a sensitive and predictive marker for antigen-specific, genetically restricted IFN-gamma production and IFN-gamma-secreting cells.. J Immunol Methods. 2001; 257(1-2):55-69. (Clone-specific: ELISA, Flow cytometry). View Reference
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Corinti S, Medaglini D, Cavani A, et al. Human dendritic cells very efficiently present a heterologous antigen expressed on the surface of recombinant gram-positive bacteria to CD4+ T lymphocytes.. J Immunol. 1999; 163(6):3029-36. (Clone-specific: ELISA). View Reference
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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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Kraan MC, Patel DD, Haringman JJ, et al. The development of clinical signs of rheumatoid synovial inflammation is associated with increased synthesis of the chemokine CXCL8 (interleukin-8).. Arthritis Res. 2001; 3(1):65-71. (Clone-specific: ELISA). 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. (Methodology: Flow cytometry, IC/FCM Block, Immunofluorescence). View Reference
566013 Rev. 1
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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.