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      V450 Mouse anti-T-bet
      V450 Mouse anti-T-bet
      Flow cytometric analysis of T-bet expression in human peripheral blood lymphocytes. Whole blood was treated with BD™ Phosflow Lyse/Fix Buffer (Cat. No. 558049) to lyse erythrocytes and fix leukocytes. The cells were then permeabilized by treatment with BD™ Phosflow Perm Buffer III (Cat. No. 558050). The cells were stained with Alexa Fluor® 700 Mouse Anti-Human CD3 (Cat. No. 557943/561027) and BD Horizon™ V450 Mouse anti-T-bet antibody (Cat. No. 561312, Right Panel) or with a BD Horizon™ V450 Mouse IgG1, κ Isotype Control (Cat. No. 560373; Left Panel). Two-color flow cytometric dot plots showing the correlated expression of T-bet (or Ig isotype control staining) versus CD3 were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
      V450 Mouse anti-T-bet
      Flow cytometric analysis of T-bet expression in human peripheral blood lymphocytes. Whole blood was treated with BD™ Phosflow Lyse/Fix Buffer (Cat. No. 558049) to lyse erythrocytes and fix leukocytes. The cells were then permeabilized by treatment with BD™ Phosflow Perm Buffer III (Cat. No. 558050). The cells were stained with Alexa Fluor® 700 Mouse Anti-Human CD3 (Cat. No. 557943/561027) and BD Horizon™ V450 Mouse anti-T-bet antibody (Cat. No. 561312, Right Panel) or with a BD Horizon™ V450 Mouse IgG1, κ Isotype Control (Cat. No. 560373; Left Panel). Two-color flow cytometric dot plots showing the correlated expression of T-bet (or Ig isotype control staining) versus CD3 were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Horizon™
      T-box expressed in T cells; TBX21; T-box 21; TBLYM
      Human (QC Testing), Mouse (Tested in Development)
      Mouse IgG1, κ
      Human T-bet Peptide
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_10611714
      Aqueous buffered solution containing protein stabilizer 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™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.
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      Recommended Assay Procedures

      Immunofluorescent staining with the O4-46 Mouse Anti-T-bet monoclonal antibody is compatible with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) and the BD Phosflow™ Permeabilization Buffers: Perm/Wash Buffer I (Cat. No. 557885); Perm Buffer II (Cat. No. 558052); Perm Buffer III (Cat. No. 558050); and Perm Buffer IV (Cat. No. 560746).

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      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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
      6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
      7. This product may be covered by US Patent No. 7,365,169.
      8. Limited Use License: The buyer (a) shall not sell or otherwise transfer this product to any third party, (b) shall use this product only for its internal, non-clinical research use, (c) shall not use this product for Commercial Purposes without a commercial license from Harvard (and if the buyer is interested in a commercial license, it should contact Harvard’s Office of Technology Development at 1350 Massachusetts Avenue, Holyoke Center, Suite 727, Cambridge, MA 02138, 617-495-3067), (d) shall use this product in compliance with all applicable laws and regulations, including, without limitation, applicable human health and animal welfare laws and regulations, (e) may transfer information or materials made through the use of this product to a scientific collaborator only if such transfer is not for any Commercial Purpose and such collaborator agrees in writing not to transfer such materials to any third party and to use such transferred materials and/or information solely for internal, non-clinical research and not for Commercial Purposes, (f) acknowledges that this product has not been approved for use in humans by the U.S. Food and Drug Administration or any other regulatory body and may not be used in humans and (g) shall indemnify, defend and hold harmless Becton Dickinson and Company and Harvard from and against all damages, losses, expenses (including reasonable attorneys’ fees), claims, demands, suits and other actions in any way arising from the buyer’s use, storage or disposal of this product.
      9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      561312 Rev. 2
      Antibody Details
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      O4-46

      The O4-46  monoclonal antibody specifically binds to human and mouse T-bet. T-bet (T-box gene expressed in T cells) is a master regulatory transcription factor that is also known as TBX21 (T-box21) and TBLYM (T-box transcription factor, expressed in lymphocytes). Human (535 amino acids; 58.3 kDa predicted molecular mass) and mouse (530 amino acids; 57.7 kDa) T-bet proteins are encoded by the human TBX21 (chromosome 17) and mouse Tbx21 (chromosome 11) genes. The human and mouse T-bet protein amino acid sequences are 88% homologous. Human and mouse T-bet proteins share a highly conserved (98% homologous amino acid sequences) T-box protein domain that is centrally located and mediates binding to DNA. T-bet is expressed by and activates transcriptional activities within hemotopoietic cells including stem cells,  NK and NKT cells and subsets of thymocytes, primed/activated CD4+ T cells, CD8+ T cells and γδ T cells, B cells, and dendritic cells. Interferon-gamma (IFN-γ), interleukin-27 (IL-27), and IL-12 act on peripheral antigen-triggered (TCR-signaling) T cells to increase T-bet expression. With respect to T helper lymphocytes, T-bet directs the differentiation of naïve CD4+ precursor T cells to become Th1-like effector and memory cells. T-bet accomplishes this by activating Th1 genetic programs (including epigenetic modifications) while repressing opposing T helper subset programs. T-bet controls the upregulated expression of the Th1 signature cytokine, IFN-γ, the IL-12Rβ2 subunit and the Runx3 transcription factor and can repress the function of other transcriptional regulators, such as GATA-3 (master regulator of Th2 development) and the expression of other cytokines including IL-2, IL-4 and IL-5.

      The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

      561312 Rev. 2
      Format Details
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      V450
      BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      V450
      Violet 405 nm
      405 nm
      450 nm
      561312 Rev.2
      Citations & References
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      View product citations for antibody "561312" on CiteAb

      Development References (10)

      1. Hibbert L, Pflanz S, De Waal Malefyt R, Kastelein RA. IL-27 and IFN-alpha signal via Stat1 and Stat3 and induce T-Bet and IL-12Rbeta2 in naive T cells. J Interferon Cytokine Res. 2003; 23(9):513-522. (Biology). View Reference
      2. Hwang ES, Hong JH, Glimcher LH. IL-2 production in developing Th1 cells is regulated by heterodimerization of RelA and T-bet and requires T-bet serine residue 508. J Exp Med. 2005; 202(9):1289-1300. (Biology). View Reference
      3. Hwang ES, Szabo SJ, Schwartzberg PL, Glimcher LH. T helper cell fate specified by kinase-mediated interaction of T-bet with GATA-3. Science. 2005; 307(5780):430-433. (Biology). View Reference
      4. Lugo-Villarino G, Maldonado-Lopez R, Possemato R, Penaranda C, Glimcher LH. T-bet is required for optimal production of IFN-gamma and antigen-specific T cell activation by dendritic cells. Proc Natl Acad Sci U S A. 2003; 100(13):7749-7754. (Biology). View Reference
      5. Peng SL, Szabo SJ, Glimcher LH. T-bet regulates IgG class switching and pathogenic autoantibody production. Proc Natl Acad Sci U S A. 2002; 99(8):5545-5550. (Biology). View Reference
      6. Peng SL. The T-box transcription factor T-bet in immunity and autoimmunity. Cell Mol Immunol. 2006; 3(2):87-95. (Biology). View Reference
      7. Szabo SJ, Kim ST, Costa GL, Zhang X, Fathman CG, Glimcher LH. A novel transcription factor, T-bet, directs Th1 lineage commitment. Cell. 2000; 100(6):655-669. (Biology). View Reference
      8. Takeda A, Hamano S, Yamanaka A, et al. Cutting edge: role of IL-27/WSX-1 signaling for induction of T-bet through activation of STAT1 during initial Th1 commitment. J Immunol. 2003; 170(10):4886-4890. (Biology). View Reference
      9. Townsend MJ, Weinmann AS, Matsuda JL, et al. T-bet regulates the terminal maturation and homeostasis of NK and Valpha14i NKT cells. Immunity. 2004; 20(4):477-494. (Biology). View Reference
      10. Zhang WX, Yang SY. Cloning and characterization of a new member of the T-box gene family. Genomics. 2000; 70(1):41-48. (Biology). View Reference
      View All (10) View Less
      561312 Rev. 2

       

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      For Research Use Only. Not for use in diagnostic or therapeutic procedures.