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Alexa Fluor® 647 Mouse anti-T-Bet
Alexa Fluor® 647 Mouse anti-T-Bet
Flow cytometric analysis of T-bet expression by 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/Wash Buffer I (Cat. No. 557885; Left Panel) or BD™ Phosflow Perm Buffer III (Cat. No. 558050; Right Panel). The cells were stained with Alexa Fluor® 647 Mouse anti-T-bet (Cat. No. 561264) and PE Mouse Anti-Human CD3 (Cat. No. 555333) antibodies. Two-color flow cytometric dot plots showing the correlated expression patterns of CD3 and T-bet were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis was performed using a BD FACSCalibur™ Flow Cytometer and FlowJo™ software.
Flow cytometric analysis of T-bet expression by 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/Wash Buffer I (Cat. No. 557885; Left Panel) or BD™ Phosflow Perm Buffer III (Cat. No. 558050; Right Panel). The cells were stained with Alexa Fluor® 647 Mouse anti-T-bet (Cat. No. 561264) and PE Mouse Anti-Human CD3 (Cat. No. 555333) antibodies. Two-color flow cytometric dot plots showing the correlated expression patterns of CD3 and T-bet were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis was performed using a BD FACSCalibur™ Flow Cytometer and FlowJo™ software.
製品詳細
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BD Pharmingen™
T-box expressed in T cells; TBX21; T-box 21; TBLYM
Human (QC Testing), Mouse (Reported)
Mouse IgG1, κ
Mouse T-bet Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_10563424
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation and Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

推奨アッセイ手順

Flow cytometry with the 4B10 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).

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
561264 Rev. 2
抗体の詳細
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4B10

The 4B10 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 hematopoietic 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.

561264 Rev. 2
フォーマットの詳細
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
561264 Rev.2
引用&参考文献
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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. (Clone-specific: Western blot). 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. (Clone-specific: Western blot). 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. (Immunogen: Western blot). 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
すべて表示する (10) 表示項目を減らす
561264 Rev. 2

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