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BV421 Mouse Anti-Human TIM-3 (CD366)
BV421 Mouse Anti-Human TIM-3 (CD366)
Multiparameter flow cytometric analysis of TIM-3 (CD366) expression on human peripheral blood cells. Human whole blood was stained with PE Mouse Anti-Human CD56 antibody (Cat. No. 555516/561903) and either BD Horizon™ BV421 Mouse IgG1 κ Isotype Control (Cat. No. 562438) or BD Horizon BV421 Mouse Anti-Human TIM-3 (CD366) antibody (Cat. No. 565562/565563). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).        Left Plots: The two-parameter flow cytometric contour plots show the correlated expression patterns of TIM-3 (CD366) expression (or Ig Isotype control staining) versus side scattered light-signals (SSC-A) distinguishing intact monocyte (intermediate SSC-A) and lymphocyte (low SCC-A) populations.        Right Plots: The two-color flow cytometric contour plots show the correlated expression of CD56 versus TIM-3 (CD366) [or Ig Isotype control staining]. Gated events with the forward and side-light scattering characteristics of intact lymphocytes are displayed.        Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multiparameter flow cytometric analysis of TIM-3 (CD366) expression on human peripheral blood cells. Human whole blood was stained with PE Mouse Anti-Human CD56 antibody (Cat. No. 555516/561903) and either BD Horizon™ BV421 Mouse IgG1 κ Isotype Control (Cat. No. 562438) or BD Horizon BV421 Mouse Anti-Human TIM-3 (CD366) antibody (Cat. No. 565562/565563). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).        Left Plots: The two-parameter flow cytometric contour plots show the correlated expression patterns of TIM-3 (CD366) expression (or Ig Isotype control staining) versus side scattered light-signals (SSC-A) distinguishing intact monocyte (intermediate SSC-A) and lymphocyte (low SCC-A) populations.        Right Plots: The two-color flow cytometric contour plots show the correlated expression of CD56 versus TIM-3 (CD366) [or Ig Isotype control staining]. Gated events with the forward and side-light scattering characteristics of intact lymphocytes are displayed.        Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
CD366; HAVCR2; TIM3; T cell immunoglobulin mucin-3; TIMD-3; KIM-3
Human (QC Testing)
Mouse IgG1, κ
Human TIM-3
Flow cytometry (Routinely Tested)
5 µl
X
84868
AB_2744369
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. 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).
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565562 Rev. 1
Antibody Details
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7D3

The 7D3 monoclonal antibody specifically binds to T cell immunoglobulin mucin 3 (TIM-3) which is also known as, CD366, or T-cell immunoglobulin and mucin domain-containing protein 3 (TIMD-3/TIMD3). CD366 is encoded by the HAVCR2 gene (Hepatitis A virus cellular receptor 2). CD366 is a type I transmembrane glycoprotein and belongs to the human TIM family (along with TIM-1 and TIM-4) within the immunoglobulin superfamily. CD366 is expressed on Th1, Tc1, Th17, Treg, NK T, and NK cells. CD366 is also expressed on dendritic cells, mast cells, monocytes, and macrophages. It is not expressed by Th2 and B cells. CD366 helps maintain peripheral immune tolerance and homeostasis. CD366 regulates macrophage activation and is a negative regulator of Th1 cell function. Crosslinking of cell surface CD366 by binding to Galectin-9 and/or phosphatidylserine appears to play an important role in either positively or negatively regulating leucocyte functions, such as cytokine production or the phagocytosis of apoptotic cells. CD366 may also be useful as an AML stem cell surface marker because it appears to be more highly expressed by AML leukemia stem cells than by normal bone marrow hematopoietic stem cells.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

565562 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV421
Violet 405 nm
407 nm
423 nm
565562 Rev.1
Citations & References
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Development References (12)

  1. Domenig C, Zheng XX, Sabatos CA, et al. Tim-3 inhibits T helper type 1-mediated auto- and alloimmune responses and promotes immunological tolerance. Nat Immunol. 2003; 4(11):1093-1101. (Biology). View Reference
  2. Freeman GJ, Casasnovas JM, Umetsu DT, DeKruyff RH. TIM genes: a family of cell surface phosphatidylserine receptors that regulate innate and adaptive immunity.. Immunol Rev. 2010; 235(1):172-89. (Biology). View Reference
  3. Hafler DA, Kuchroo V. TIMs: Central regulators of immune responses. J Exp Med. 2008; 205:2699-2701. (Biology). View Reference
  4. Jan M, Chao MP, Cha AC, et al. Prospective separation of normal and leukemic stem cells based on differential expression of TIM3, a human acute myeloid leukemia stem cell marker. Proc Natl Acad Sci U S A. 2011; 108(12):5009-5014. (Biology). View Reference
  5. Khademi M, Illes Z, Gielen AW, et al. T Cell Ig- and mucin-domain-containing molecule-3 (TIM-3) and TIM-1 molecules are differentially expressed on human Th1 and Th2 cells and in cerebrospinal fluid-derived mononuclear cells in multiple sclerosis. J Immunol. 2004; 172(11):7169-7176. (Biology). View Reference
  6. Lee J, Su EW, Zhu C, et al. Phosphotyrosine-dependent coupling of Tim-3 to T-cell receptor signaling pathways. Mol Cell Biol. 2011; 31(19):3963-3974. (Biology). View Reference
  7. Lee JS, Park MJ, Park S, Lee ES. Differential expression of T cell immunoglobulin- and mucin-domain-containing molecule-3 (TIM-3) according to activity of Behcet's disease. Br J Dermatol. 2012; 65(3):220-222. (Biology). View Reference
  8. Moorman JP, Wang JM, Zhang Y, et al. Tim-3 pathway controls regulatory and effector T cell balance during hepatitis C virus infection. J Immunol. 2012; 189(2):755-766. (Biology). View Reference
  9. Ndhlovu LC, Lopez-Verges S, Barbour JD, et al. Tim-3 marks human natural killer cell maturation and suppresses cell-mediated cytotoxicity. Blood. 2012; 119(16):3734-3743. (Biology). View Reference
  10. Rodriguez-Manzanet R, DeKruyff R, Kuchroo VK, Umetsu DT. The costimulatory role of TIM molecules. Immunol Rev. 2009; 229(1):259-270. (Biology). View Reference
  11. Wang F, Wan L, Zhang C, Zheng X, Li J, Chen ZK. Tim-3-Galectin-9 pathway involves the suppression induced by CD4+CD25+ regulatory T cells. Immunobiology. 2009; 214(5):342-349. (Biology). View Reference
  12. van de Weyer PS, Muehlfeit M, Klose C, Bonventre JV, Walz G, Kuehn EW. A highly conserved tyrosine of Tim-3 is phosphorylated upon stimulation by its ligand galectin-9. Biochem Biophys Res Commun. 2006; 351(2):571-576. (Biology). View Reference
View All (12) View Less
565562 Rev. 1

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