Recombinant Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR27094-16] to T-bet / Tbx21
- Suitable for: ICC/IF, Flow Cyt (Intra), IHC-P, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-T-bet / Tbx21 antibody [EPR27094-16]
See all T-bet / Tbx21 primary antibodies -
Description
Rabbit monoclonal [EPR27094-16] to T-bet / Tbx21 -
Host species
Rabbit -
Specificity
ICC/IF: Unsuitable for mouse samples
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Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra), IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR27094-16 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab307193 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC/IF |
1/500.
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Flow Cyt (Intra) |
1/500.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Predicted molecular weight: 58 kDa.
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Notes |
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ICC/IF
1/500. |
Flow Cyt (Intra)
1/500. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 58 kDa. |
Target
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Function
Transcription factor that controls the expression of the TH1 cytokine, interferon-gamma. Initiates TH1 lineage development from naive TH precursor cells both by activating TH1 genetic programs and by repressing the opposing TH2 programs. -
Tissue specificity
T-cell specific. -
Involvement in disease
Genetic variations in TBX21 are associated with susceptibility to asthma with nasal polyps and aspirin intolerance (ANPAI) [MIM:208550]. A condition consisting of asthma, aspirin sensitivity and nasal polyposis. Nasal polyposis is due to chronic inflammation of the paranasal sinus mucosa, leading to protrusion of edematous polyps into the nasal cavities. -
Sequence similarities
Contains 1 T-box DNA-binding domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 30009 Human
- Entrez Gene: 57765 Mouse
- Entrez Gene: 303496 Rat
- Omim: 604895 Human
- SwissProt: Q9UL17 Human
- SwissProt: Q9JKD8 Mouse
- Unigene: 272409 Human
- Unigene: 94519 Mouse
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Alternative names
- T bet antibody
- T box 21 antibody
- T box expressed in T cells antibody
see all
Images
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All lanes : Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution
Lane 1 : No-GFP-CD16. NK-92 (human malignant non-Hodgkin's 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 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 58 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?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.
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Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution + mouse spleen tissue lysate 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 58 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?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.
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Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution + rat spleen tissue lysate 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 58 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?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.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (1.044 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma (T cell) tissue labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (1.044 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (1.044 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (1.044 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded mouse Burkitt's lymphoma tissue labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (1.044 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (1.044 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-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 dilution (1.044 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (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). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml) (Red). Nuclear counterstain was DAPI (Blue).Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/ml).
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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 compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
Live cells were stained with anti-CD56 conjµgated to BV421.
Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or 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 (ab172730) / Left isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, 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.
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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 labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (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, ab150081) at 1/2000 dilution was used as the secondary antibody.
Negative control: K562. -
Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized mouse splenocytes labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, 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.
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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 (ab172730) / Left isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, 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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab307193 has been referenced in 2 publications.
- Zhang H et al. TFAP2C exacerbates psoriasis-like inflammation by promoting Th17 and Th1 cells activation through regulating TEAD4 transcription. Allergol Immunopathol (Madr) 51:124-134 (2023). PubMed: 37169570
- Li S et al. HK3 stimulates immune cell infiltration to promote glioma deterioration. Cancer Cell Int 23:227 (2023). PubMed: 37779195