Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)

Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) is a rabbit monoclonal antibody that is used to detect T-bet / Tbx21 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	Flow Cyt (Intra)	ICC/IF	IHC-P	WB
Human	Tested	Tested	Tested	Tested
Mouse	Tested	Not recommended	Tested	Tested
Rat	Expected	Expected	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Target data

See full target information TBX21

Function

Lineage-defining transcription factor which initiates Th1 lineage development from naive Th precursor cells both by activating Th1 genetic programs and by repressing the opposing Th2 and Th17 genetic programs (PubMed:10761931). Activates transcription of a set of genes important for Th1 cell function, including those encoding IFN-gamma and the chemokine receptor CXCR3. Induces permissive chromatin accessibilty and CpG methylation in IFNG (PubMed:33296702). Activates IFNG and CXCR3 genes in part by recruiting chromatin remodeling complexes including KDM6B, a SMARCA4-containing SWI/SNF-complex, and an H3K4me2-methyltransferase complex to their promoters and all of these complexes serve to establish a more permissive chromatin state conducive with transcriptional activation (By similarity). Can activate Th1 genes also via recruitment of Mediator complex and P-TEFb (composed of CDK9 and CCNT1/cyclin-T1) in the form of the super elongation complex (SEC) to super-enhancers and associated genes in activated Th1 cells (PubMed:27292648). Inhibits the Th17 cell lineage commitment by blocking RUNX1-mediated transactivation of Th17 cell-specific transcriptinal regulator RORC. Inhibits the Th2 cell lineage commitment by suppressing the production of Th2 cytokines, such as IL-4, IL-5, and IL- 13, via repression of transcriptional regulators GATA3 and NFATC2. Protects Th1 cells from amplifying aberrant type-I IFN response in an IFN-gamma abundant microenvironment by acting as a repressor of type-I IFN transcription factors and type-I IFN-stimulated genes. Acts as a regulator of antiviral B-cell responses; controls chronic viral infection by promoting the antiviral antibody IgG2a isotype switching and via regulation of a broad antiviral gene expression program (By similarity). Required for the correct development of natural killer (NK) and mucosal-associated invariant T (MAIT) cells (PubMed:33296702).

Alternative names

TBET, TBLYM, TBX21, T-box transcription factor TBX21, T-box protein 21, T-cell-specific T-box transcription factor T-bet, Transcription factor TBLYM

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR27094-16
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), in Human, Mouse, Rat samples.

What is the molecular weight of T-bet / Tbx21?

Anti-T-bet / Tbx21 [EPR27094-16] (ab307193) specifically detects a band for T-bet / Tbx21 (UniProt: Q9UL17) at a molecular weight of 58kDa.

Recommended positive controls

WB: No-GFP-CD16. NK-92, K-562, Mouse spleen and Rat spleen lysates.IHC-P: Human tonsil, Human non-Hodgkin's lymphoma, Mouse spleen, Rat spleen, Mouse Burkitt's lymphoma and Human cerebrum tissues.ICC/IF: No-GFP-CD16.NK-92 cells. Flow Cyt: Human peripheral blood mononuclear, Human peripheral blood mononuclear, K562, Mouse peripheral blood mononuclear and Mouse splenocytes cells.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.Other related products

We have a range of other formats of antibody clone [EPR27094-16] also available for your convenience:
ab307193, Carrier free - ab307194, Alexa Fluor® 488 - ab315424, Alexa Fluor® 594 - ab315425, Alexa Fluor® 647 - ab315957

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

T-bet also known as Tbx21 is a T-box transcription factor that plays an important role in regulating immune responses. T-bet has a molecular mass of approximately 58 kDa. It is extensively expressed in T-helper 1 (Th1) cells and natural killer (NK) cells facilitating these cells' development and function. T-bet guides the transcriptional activities necessary for cellular differentiation and immune response by binding to specific DNA sequences. Its expression is tightly controlled reflecting its central role in immune regulation. "9d bet" is also referenced in scientific research although its specific context requires precise understanding within a given study.

Biological function summary

This transcription factor is pivotal for manufacturing interferon-gamma (IFN-γ) initiating and sustaining Th1 cell immunity. T-bet doesn't act alone; it functions as part of larger protein complexes interacting with other transcription factors and cofactors to exert its effects. The capacity to influence IFN-γ production makes T-bet a significant player in immune responses helping coordinate responses against intracellular pathogens. Moreover the expression of T-bet is not limited solely to T cells but also impacts other immune cells like CD8+ T cells and B cells. Anti-bet or anti-T antibodies frequently target T-bet in research to explore immune system intricacies better.

Pathways

T-bet integrates into the immune signaling network through pathways such as the JAK-STAT pathway and the Th1 differentiation pathway. It activates transcription of the IFNG gene working closely with related proteins like STAT4. T-bet's influence extends as it cooperates with STAT1 enabling a feed-forward loop that amplifies and stabilizes Th1 responses. "Bet t products" could refer to those derived from such transcriptional activation. Additionally T-bet involvement goes beyond transcriptional regulation affecting cytokine transport and secretion processes vital for an effective immune response.

Associated diseases and disorders

Researchers have linked T-bet to autoimmune diseases and infectious diseases. Aberrant expression of T-bet can lead to enhanced Th1 responses intensifying autoimmune conditions such as multiple sclerosis (MS). Its role in diseases connects it with NF-κB pathways and other transcription factors influencing inflammatory processes. Furthermore "anti bet" studies often involve understanding T-bet's role in infectious diseases seeking to manipulate its function to adjust immune responses in chronic infections. Studies targeting its pathways or interactions with disease-specific proteins aim to develop therapeutic strategies turning an element of dysregulation into a novel treatment approach.

Product promise

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15 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control: No staining on human cerebrum. The section was incubated with ab307193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Immunohistochemical analysis of paraffin-embedded Mouse Burkitt's lymphoma tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on mouse Burkitt's lymphoma. The section was incubated with ab307193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on rat spleen. The section was incubated with ab307193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on mouse spleen. The section was incubated with ab307193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin's tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on human non-Hodgkin's lymphoma (T cell). The section was incubated with ab307193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on human tonsil. The section was incubated with ab307193 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The lysate was freshly made and used for Western Blotting immediately to minimize protein degradation.

Exposure time: 180 seconds
All lanes: Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution
All lanes: Mouse spleen tissue lysate 20 μg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 60 kDa
Exposure time: 180s
Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse splenocytes cells labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Live cells were stained with anti-CD3 conjugated to Alexa Fluor® 647. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab307193.
Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) cells labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: CD19+ population. Live cells were stained with anti-CD19 conjugated to Alexa Fluor® 647. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab307193.
Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The lysate was freshly made and used for Western Blotting immediately to minimize protein degradation.

The identity of the bands below 50 kDa are unknown.
Exposure time: 103 seconds
All lanes: Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution
All lanes: Rat spleen tissue lysate 20 μg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 60 kDa
Exposure time: 103s
Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: K-562

In lane 1, the lysate was freshly made and used for Western Blotting immediately to minimize protein degradation.

The identity of the bands below 50 kDa are unknown.
Exposure time: 15 seconds
All lanes: Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution
Lane 1: No-GFP-CD16.NK-92 (human malignant non-Hodgkins lymphoma natural killer cell) whole cell lysate 20 μg
Lane 2: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 20 μg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 60 kDa
Exposure time: 15s
Immunocytochemistry/ Immunofluorescence - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer (NK-92®) cell with no GFP tag) cells labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2µg/ml dilution (Green). Confocal image showing nuclear staining in No-GFP-CD16.NK-92 cell line. Negative control: K562. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2µg/ml dilution.
Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse peripheral blood mononuclear cell (PBMC) cells labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: CD19+ population. Live cells were stained with anti-CD19 conjugated to Alexa Fluor® 647. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab307193.
Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K562 (human chronic myelogenous leukemia lymphoblast, Left) / No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag, Right) cells labelling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: K562.
Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) cells labelling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Live cells were stained with anti-CD56 conjugated to BV421. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab307193.