Skip to main content Skip to navigation
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.
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
Down Arrow Up Arrow


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.

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.
569318 Rev. 1
Antibody Details
Down Arrow Up Arrow
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
Down Arrow Up Arrow
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
Down Arrow Up Arrow

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

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.