Clone:
REA102
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC, MC
Alternative names:
TBX21, T-PET, TBLYM, TBT1

Extended validation for T-bet Antibody, anti-human/mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA102
4B10++
O4-46++
Cells were incubated with an excess of purified unconjugated T-bet (REA102) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with ViobilityTM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with ViobilityTM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with ViobilityTM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with ViobilityTM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with ViobilityTM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for T-bet Antibody, anti-human/mouse, REAfinity™

Overview

The monoclonal antibody REA102 recognizes human and mouse T-bet, a Tʜ1-specific T-box 1 transcription factor. T-bet is considered to be an important regulator of Tʜ1 lymphoid development and is involved in class-switch recombination in B cells. It is widely expressed in hematopoietic cells such as B cells, T cell subsets, NK and NKT cells, and dendritic cells. Expression of T-bet is induced by Tʜ1 cytokine IFN-γ and T-bet further regulates the expression of cytokines including IFN-γ, IL-12Rβ, and IL-2.
Additional information: Clone REA102 displays negligible binding to Fc receptors.

Alternative names

TBX21, T-PET, TBLYM, TBT1

Detailed product information

Technical specifications

CloneREA102
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse
AntigenT-bet
Alternative names of antigenTBX21, T-PET, TBLYM, TBT1
Molecular mass of antigen [kDa]58
Entrez Gene ID30009
RRIDAB_2811373, AB_2784465, AB_2784464, AB_2784467, AB_2784466, AB_2801868, AB_2811376

Resources for T-bet Antibody, anti-human/mouse, REAfinity™

Certificates

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References for T-bet Antibody, anti-human/mouse, REAfinity™

Publications

  1. Szabo, S. J. et al. (2002) Distinct effects of T-bet in Th1 lineage commitment and IFN-gamma production in CD4 and CD8 T cells. Science 295: 338
  2. Lugo-Villarino, G. et al. (2003) T-bet is required for optimal production of IFN-γ and antigen-specific T cell activation by dendritic cells. Proc. Natl. Acad. Sci. U.S.A. 100: 7749-7754
  3. Peng, S. L. et al. (2002) T-bet regulates IgG class switching and pathogenic autoantibody production. Proc. Natl. Acad. Sci. U.S.A. 99: 5545-5550
  4. Szabo, S. J. et al. (2000) A novel transcription factor, T-bet, directs Tʜ1 lineage commitment. Cell 100(6): 655-669
  5. Qian, L. et al. (2014) Alkylglycerols modulate the proliferation and differentiation of non-specific agonist and specific antigen-stimulated splenic lymphocytes. PLoS One 9(4): e96207

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