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      Purified NA/LE Rat Anti-Mouse CD366 (TIM-3)
      Purified NA/LE Rat Anti-Mouse CD366 (TIM-3)
      Flow cytometric analysis of Recombinant Mouse TIM-3 protein binding blockade to phosphatidylserine expressed on apoptotic Mouse thymocytes by Purified NA/LE Rat Anti-Mouse CD366 (TIM-3) antibody. BALB/c Mouse thymocytes were cultured in the presence of 1 µM of Dexamethasone for 5 h (37°C) to induce apoptosis. Apoptotic thymocytes (1 x 10^6 cells/0.1 ml final volume) were mixed with Recombinant Mouse TIM-3 Fc Chimera Protein (5 µg test dose; R&D Systems, Cat. No. 1529-TM-050) preincubated (30 min 4°C) with either Purified NA/LE Rat Anti-Mouse CD366 (TIM-3) antibody (Cat. No. 569318; Solid Line) or Purified NA/LE Rat IgG2a Isotype Control (Cat. No. 554687; Dotted Line) at the indicated test doses. The cells were washed and secondarily stained with Biotin Goat Anti-Human IgG, Fc fragment specific (Biotin Anti-Human IgG Fc; Jackson ImmunoResearch, Cat. No. 109-065-190) followed by PE Streptavidin (Cat. No. 554061). The graphs showing the percentage of cells bound with Recombinant TIM-3+ (% Recombinant TIM-3+ Cells) after preincubation with TIM-3-specific Blocking Antibody (or Ig Isotype Control) were derived from gated events with the forward and side light-scatter characteristics of apoptotic thymocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Purified NA/LE Rat Anti-Mouse CD366 (TIM-3)
      Flow cytometric analysis of Recombinant Mouse TIM-3 protein binding blockade to phosphatidylserine expressed on apoptotic Mouse thymocytes by Purified NA/LE Rat Anti-Mouse CD366 (TIM-3) antibody. BALB/c Mouse thymocytes were cultured in the presence of 1 µM of Dexamethasone for 5 h (37°C) to induce apoptosis. Apoptotic thymocytes (1 x 10^6 cells/0.1 ml final volume) were mixed with Recombinant Mouse TIM-3 Fc Chimera Protein (5 µg test dose; R&D Systems, Cat. No. 1529-TM-050) preincubated (30 min 4°C) with either Purified NA/LE Rat Anti-Mouse CD366 (TIM-3) antibody (Cat. No. 569318; Solid Line) or Purified NA/LE Rat IgG2a Isotype Control (Cat. No. 554687; Dotted Line) at the indicated test doses. The cells were washed and secondarily stained with Biotin Goat Anti-Human IgG, Fc fragment specific (Biotin Anti-Human IgG Fc; Jackson ImmunoResearch, Cat. No. 109-065-190) followed by PE Streptavidin (Cat. No. 554061). The graphs showing the percentage of cells bound with Recombinant TIM-3+ (% Recombinant TIM-3+ Cells) after preincubation with TIM-3-specific Blocking Antibody (or Ig Isotype Control) were derived from gated events with the forward and side light-scatter characteristics of apoptotic thymocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      Cd366; TIM-3; Tim3; TIMD-3; Timd3; HAVcr-2; Havcr2
      Mouse (QC Testing)
      Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
      Mouse TIM-3 Recombinant Protein
      Blocking, Flow cytometry (Tested During Development)
      1.0 mg/ml
      171285
      No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.
      RUO


      Preparation And Storage

      Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      4. An isotype control should be used at the same concentration as the antibody of interest.
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      569318 Rev. 1
      Antibody Details
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      RMT3-23

      The RMT3-23 monoclonal antibody specifically recognizes CD366 which is also known as TIM-3 (T-cell immunoglobulin and mucin domain-containing 3,  T-cell immunoglobulin mucin receptor 3, or T-cell membrane protein 3). CD366 (TIM-3) is ~31 kDa type I transmembrane glycoprotein encoded by Havcr2 (Hepatitis A virus cellular receptor 2) that belongs to the TIM family within the immunoglobulin superfamily (IgSF). CD366 (TIM-3) is comprised of one Ig-like V-type domain followed by a serine/threonine-rich mucin stalk region in its extracellular region, a transmembrane segment, and a tyrosine phosphorylation motif in its cytoplasmic tail. CD366 (TIM-3) expression is upregulated on subpopulations of activated myeloid cells including macrophages, monocytes, dendritic cells (DC), microglia, mast cells as well as on Type-1 CD4+ (Th1-like) T cells, cytotoxic CD8+ T cells, regulatory T cells (Treg), and natural killer (NK) cells. CD366 (TIM-3) functions as an inhibitory receptor that helps maintain immunological homeostasis and self-tolerance. It may also serve an immune checkpoint molecule that inhibits antitumor immunity and promotes T cell exhaustion. Crosslinking of cell surface CD366 (TIM-3) by Galectin-9 binding downregulates Th1-like and CD8+ T cell responses and can promote Treg or myeloid-derived suppressor cells. CD366 (TIM-3) enables DC to bind phosphatidyl serine expressed by apoptotic cells, phagocytize these cells to suppress inflammation and promote antigen cross-presentation. CD366 (TIM-3) can also bind to high mobility group protein B1 (HMGB1) and inhibit stimulation of the immune response to nucleic acids released by dying tumor cells. RMT3-23 reportedly recognizes CD366 (TIM-3) derived from either BALB/c or C57BL/6 mice that have different Havcr2 polymorphisms.

      569318 Rev. 1
      Format Details
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      NA/LE
      NA/LE refers to the culture and purification methods and buffer used to produce purified antibodies with no azide and low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.NA/LE are perfectly suited to be used in culture or in vivo (for nonhuman studies) for functional assays — blocking, neutralizing, activation or depletion — where the presence of azide may damage cells or exogenous endotoxin may signal or activate cells.
      NA/LE
      569318 Rev.1
      Citations & References
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      View product citations for antibody "569318" on CiteAb

      Development References (11)

      1. Anderson AC, Joller N, Kuchroo VK. Lag-3, Tim-3, and TIGIT: Co-inhibitory Receptors with Specialized Functions in Immune Regulation.. Immunity. 2016; 44(5):989-1004. (Biology). View Reference
      2. Anderson AC, Xiao S, Kuchroo VK. Tim protein structures reveal a unique face for ligand binding.. Immunity. 2007; 26(3):273-5. (Biology). View Reference
      3. Chiba S, Baghdadi M, Akiba H, et al. Tumor-infiltrating DCs suppress nucleic acid-mediated innate immune responses through interactions between the receptor TIM-3 and the alarmin HMGB1.. Nat Immunol. 2012; 13(9):832-42. (Biology). View Reference
      4. Nakae S, Iikura M, Suto H, et al. TIM-1 and TIM-3 enhancement of Th2 cytokine production by mast cells.. Blood. 2007; 110(7):2565-8. (Clone-specific: Flow cytometry). View Reference
      5. Nakayama M, Akiba H, Takeda K, et al. Tim-3 mediates phagocytosis of apoptotic cells and cross-presentation. Blood. 2009; 113(16):3821-3830. (Clone-specific). View Reference
      6. Ngiow SF, von Scheidt B, Akiba H, Yagita H, Teng MW, Smyth MJ. Anti-TIM3 antibody promotes T cell IFN-γ-mediated antitumor immunity and suppresses established tumors.. Cancer Res. 2011; 71(10):3540-51. (Clone-specific: Flow cytometry). View Reference
      7. Oikawa T1, Kamimura Y, Akiba H, et al. Preferential involvement of Tim-3 in the regulation of hepatic CD8+ T cells in murine acute graft-versus-host disease.. J Immunol. 2006; 177(7):4281-4287. (Immunogen: Flow cytometry, Immunohistochemistry, In vivo exacerbation). View Reference
      8. Sabatos-Peyton CA, Nevin J, Brock A, et al. Blockade of Tim-3 binding to phosphatidylserine and CEACAM1 is a shared feature of anti-Tim-3 antibodies that have functional efficacy.. Oncoimmunology. 7(2):e1385690. (Clone-specific: Blocking, Flow cytometry). View Reference
      9. Sakuishi K, Ngiow SF, Sullivan JM, et al. TIM3+FOXP3+ regulatory T cells are tissue-specific promoters of T-cell dysfunction in cancer.. Oncoimmunology. 2013; 2(4):e23849. (Clone-specific: Flow cytometry). View Reference
      10. Veenstra RG, Taylor PA, Zhou Q, et al. Contrasting acute graft-versus-host disease effects of Tim-3/galectin-9 pathway blockade dependent upon the presence of donor regulatory T cells.. Blood. 2012; 120(3):682-90. (Biology). View Reference
      11. Zhu C, Anderson AC, Schubart A, et al. The Tim-3 ligand galectin-9 negatively regulates T helper type 1 immunity.. Nat Immunol. 2005; 6(12):1245-52. (Methodology). View Reference
      View All (11) View Less
      569318 Rev. 1

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      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.