Multiparameter flow cytometric analysis of Galectin-9 expression on human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human Galectin-9 antibody (Cat. No. 565890/565892; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of Galectin-9 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
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Multiparameter flow cytometric analysis of Galectin-9 expression on human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human Galectin-9 antibody (Cat. No. 565890/565892; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of Galectin-9 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Preparation and Storage
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.
関連製品
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The 9M1-3 monoclonal antibody specifically binds to Galectin-9 (Gal-9) which is encoded by LGALS9 (lectin, galactoside-binding, soluble, 9). Galectin-9 is differentially expressed within or on the surface of various cell types including lymphocytes, macrophages, astrocytes, mast cells, eosinophils, fibroblasts, and endothelial cells. Its expression can be induced by cells in response to proinflammatory cytokines or infection by certain viruses. Galectin-9 is an ~ 50 kDa S-type lectin that has two carbohydrate recognition domains (CRDs). It binds to β-galactosides and can serve as a ligand for TIM-3 (CD366). Cell-surface or released forms of Galectin-9 can play multiple roles in innate and adaptive immune responses including the activation of macrophage cytokine secretion, promotion of dendritic cell maturation and regulatory T cell expansion, and negative regulation of Th1, Th17, NK, and cytotoxic T cells. Galectin-9 may contribute to the survival and growth of some tumors by suppressing anti-tumor immune responses. The 9M1-3 antibody reportedly binds to the CRD2 domain of Galectin-9 and can block its binding to TIM-3.
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Development References (7)
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Anderson AC, Anderson DE. TIM-3 in autoimmunity.. Curr Opin Immunol. 2006; 18(6):665-9. (Biology). View Reference
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Cedeno-Laurent F, Dimitroff CJ. Galectins and their ligands: negative regulators of anti-tumor immunity.. Glycoconj J. 2012; 29(8-9):619-25. (Biology). View Reference
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Dai SY, Nakagawa R, Itoh A, et al. Galectin-9 induces maturation of human monocyte-derived dendritic cells.. J Immunol. 2005; 175(5):2974-81. (Biology). View Reference
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Elahi S, Niki T, Hirashima M, Horton H. Galectin-9 binding to Tim-3 renders activated human CD4+ T cells less susceptible to HIV-1 infection.. Blood. 2012; 119(18):4192-204. (Clone-specific: Flow cytometry). View Reference
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Gieseke F, Kruchen A, Tzaribachev N, Bentzien F, Dominici M, Müller I. Proinflammatory stimuli induce galectin-9 in human mesenchymal stromal cells to suppress T-cell proliferation.. Eur J Immunol. 2013; 43(10):2741-9. (Clone-specific: Flow cytometry). View Reference
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Klibi J, Niki T, Riedel A, et al. Blood diffusion and Th1-suppressive effects of galectin-9-containing exosomes released by Epstein-Barr virus-infected nasopharyngeal carcinoma cells.. Blood. 2009; 113(9):1957-66. (Immunogen: Blocking, Flow cytometry, Functional assay, Inhibition). View Reference
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Qiu Y, Chen J, Liao H, et al. Tim-3-expressing CD4+ and CD8+ T cells in human tuberculosis (TB) exhibit polarized effector memory phenotypes and stronger anti-TB effector functions.. PLoS Pathog. 2012; 8(11):e1002984. (Clone-specific: Flow cytometry). View Reference
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