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      PE Mouse Anti-Mouse CD366 (TIM-3)
      PE Mouse Anti-Mouse CD366 (TIM-3)
      Flow cytometric analysis of CD366 (TIM-3) expressed on activated mouse splenocytes. BALB/c splenic leucocytes were cultured for 4 days in the presence of plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 μg/ml for coating), soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 2 μg/ml), and Purified NA/LE Rat Anti-Mouse IL-4 (Cat. No. 554432) antibodies. The cells were harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and stained with Alexa Fluor® 647 Anti-Mouse CD8a antibody (Cat. No. 557682) and (at 0.25 μg per test) either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Mouse CD366 (TIM-3) antibody (Cat. No. 566346; Right Plot). After staining and washing, the cells were suspended in BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816). A contour plot showing the correlated expression of CD8a versus CD366 (TIM-3) [or Ig Isotype control staining] was derived from 7-AAD-negative gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
      PE Mouse Anti-Mouse CD366 (TIM-3)
      Flow cytometric analysis of CD366 (TIM-3) expressed on activated mouse splenocytes. BALB/c splenic leucocytes were cultured for 4 days in the presence of plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 μg/ml for coating), soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 2 μg/ml), and Purified NA/LE Rat Anti-Mouse IL-4 (Cat. No. 554432) antibodies. The cells were harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and stained with Alexa Fluor® 647 Anti-Mouse CD8a antibody (Cat. No. 557682) and (at 0.25 μg per test) either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Mouse CD366 (TIM-3) antibody (Cat. No. 566346; Right Plot). After staining and washing, the cells were suspended in BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816). A contour plot showing the correlated expression of CD8a versus CD366 (TIM-3) [or Ig Isotype control staining] was derived from 7-AAD-negative gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      Cd366; T-cell membrane protein; Tim3; TIMD-3; Timd3; HAVcr-2; Havcr2;
      Mouse (QC Testing)
      Mouse IgG1, κ
      Mouse TIM-3 Recombinant Protein
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      AB_2739702
      Aqueous buffered solution containing ≤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.
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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. 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.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
      6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      566346 Rev. 1
      Antibody Details
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      5D12/TIM-3

      The 5D12 monoclonal antibody specifically recognizes CD366 which is also known as TIM-3 (T-cell immunoglobulin and mucin domain-containing 3) or T-cell membrane protein 3. TIM-3 is encoded by Havcr2 (hepatitis A virus cellular receptor 2). TIM-3 is a type I transmembrane glycoprotein that belongs to the human TIM family within the immunoglobulin superfamily, having one Ig-like V-type domain in its extracellular region. TIM-3 is expressed on activated monocytes, macrophages, dendritic cells, microglia, and mast cells. It is also expressed on type-1 CD4+ (Th1-like) T cells, CD8+ T cytotoxic cells, regulatory T cells (Treg), and natural killer (NK) cells. TIM-3 functions as an inhibitory receptor which helps cells maintain immunological homeostasis and self tolerance. Crosslinking of cell surface TIM-3 by Galectin-9 binding downregulates Th1-like and CD8+ T cell responses and can promote Treg or myeloid-derived suppressor cells. TIM-3 enables dendritic cells to bind phosphatidyl serine expressed by apoptotic cells and to phagocytize these cells to quell potential inflammation. TIM-3 also binds to the alarmin HMGB1 thereby preventing Toll-like receptor (TLR) by released tumor cell DNA.

      566346 Rev. 1
      Format Details
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      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.
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      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      566346 Rev.1
      Citations & References
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      Development References (4)

      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. 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. (Biology). View Reference
      3. Phong BL, Avery L, Sumpter TL, et al. Tim-3 enhances FcεRI-proximal signaling to modulate mast cell activation.. J Exp Med. 2015; 212(13):2289-304. (Clone-specific: Flow cytometry). View Reference
      4. 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. (Clone-specific: Flow cytometry). View Reference
      View All (4) View Less
      566346 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.

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