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Anti-TCF-4 antibody [NCI-R159-6] ab217668 is a rabbit monoclonal antibody that is used in TCF-4 western blotting, IHC and flow cytometry. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone NCI-R159-6 is the most widely used clone for TCF-4 on the market and is cited in >30 publications
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCF-4 antibody [NCI-R159-6] (AB217668), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCF-4 antibody [NCI-R159-6] (AB217668), expandable thumbnail
  • Western blot - Anti-TCF-4 antibody [NCI-R159-6] (AB217668), expandable thumbnail
  • ChIP - Anti-TCF-4 antibody [NCI-R159-6] (AB217668), expandable thumbnail
  • Western blot - Anti-TCF-4 antibody [NCI-R159-6] (AB217668), 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
ChIPIHC-PWBFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Mouse
Not recommended
Not recommended
Tested
Not recommended

Tested
Tested

Species

Human

Dilution info

2.5 µg/mL

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

Fixative: 4% Formalin
Antigen retrieval: Low pH retrieval solution

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

Fixative: 4% Formalin
Antigen retrieval: Low pH retrieval solution

Tested
Tested

Species

Human

Dilution info

1/10000

Notes

-

Species

Mouse

Dilution info

1/1000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

-

Target data

Function

Transcription factor that binds to the immunoglobulin enhancer Mu-E5/KE5-motif. Involved in the initiation of neuronal differentiation. Activates transcription by binding to the E box (5'-CANNTG-3'). Binds to the E-box present in the somatostatin receptor 2 initiator element (SSTR2-INR) to activate transcription (By similarity). Preferentially binds to either 5'-ACANNTGT-3' or 5'-CCANNTGG-3'.

Alternative names

BHLHB19, ITF2, SEF2, ITF2, BHLHB19, TCF4, SEF2, Transcription factor 4, TCF-4, Class B basic helix-loop-helix protein 19, Immunoglobulin transcription factor 2, SL3-3 enhancer factor 2, bHLHb19, ITF-2, SEF-2

Recommended products

Anti-TCF-4 antibody [NCI-R159-6] ab217668 is a rabbit monoclonal antibody that is used in TCF-4 western blotting, IHC and flow cytometry. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone NCI-R159-6 is the most widely used clone for TCF-4 on the market and is cited in >30 publications
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

NCI-R159-6

Purification technique

Affinity purification Protein A

Specificity

Mouse specificity in WB only.
Weak human specificity in IHC.

Concentration
Loading...

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

For detailed protocol using this antibody for IHC, ChIP, and Flow Cyt, please refer to the following paper:

Ceribelli et al. A Druggable TCF4- and BRD4-Dependent Transcriptional Network Sustains Malignancy in Blastic Plasmacytoid Dendritic Cell Neoplasm, PMID: 27846392

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

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

The transcription factor 4 (TCF-4) also known as TCF7L2 is an important regulator of transcriptional activity. This protein weighs approximately 68 kDa and is widely expressed in multiple tissues including the pancreas intestine and brain. Through its function as a transcription factor TCF-4 binds to specific DNA sequences influencing the expression of target genes. The regulation it provides is essential for various biological processes especially during developmental stages.

Biological function summary

TCF-4 plays an important role in Wnt signaling a pathway critical for cell growth and differentiation. It partners with β-catenin to form a functional complex enabling it to activate gene expression. This interaction influences cellular processes such as cell cycle regulation fate determination and maintenance of stem cell populations. As such TCF-4 serves an important role in regulating a balance between proliferation and differentiation in developing tissues.

Pathways

TCF-4 is an important component within the canonical Wnt/β-catenin signaling pathway. This pathway is pivotal in embryogenesis and is also involved in the control of cell communication processes. TCF-4 interacts with other pathway members such as the APC protein which are essential for stabilizing β-catenin enabling successful transcriptional regulation. Its activity connects it chemically to the TCF/LEF family of proteins which all participate actively in gene transcription related to Wnt signaling.

Associated diseases and disorders

TCF-4 has been implicated in type 2 diabetes and certain forms of cancer. Research shows that mutations in the TCF7L2 gene alter its function leading to increased susceptibility to type 2 diabetes emphasizing its role in glucose metabolism. Moreover dysregulation of the Wnt signaling pathway in which TCF-4 is an actor has been associated with cancer pathogenesis including colorectal cancer. TCF-4 connects closely with β-catenin and APC in these conditions highlighting its involvement in tumorigenic processes when its regulation fails.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    Immunohistochemical analysis of 4% Formalin fixed Blastic plasmacytoid dendritic cell neoplasm (BPDCN) cell pellets after selection and induction of shRNA expression for 1 day, labeling TCF-4 with ab217668 at 1/100 dilution. Universal DAB Detection Kit was used for detection of IHC staining on an automated system.

    The data was provided by our collaborator Dr. Louis M. Staudt, and published in Cancer Cell 30, 764-778, 2016 (PMID: 27846392). Several TCF-4 shRNAs were used. This IHC image shows data for shRNA TCF4 #2.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    Immunohistochemical analysis of 4% Formalin fixed human tonsil labeling TCF-4 with ab217668 at 1/100 dilution. Universal DAB Detection Kit was used for detection of IHC staining on an automated system.

    pDCs: plasmacytoid dendritic cells.

    GC: Germinal Center.

    The data was provided by our collaborator Dr. Louis M. Staudt, and published in Cancer Cell 30, 764-778, 2016 (PMID: 27846392).

  • Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    The data was provided by the collaborator Dr. Louis M. Staudt, NCI, NIH.

    All lanes: Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668) at 1/10000 dilution

    Lane 1: Nuclear extracts of Cal-1 cells (Human plasmacytoid dendritic cell line) infected with control TCF4 shRNA at 20 µg

    Lane 2: Nuclear extracts of Cal-1 cells infected with TCF4 shRNA at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated

    Predicted band size: 71 kDa

    Observed band size: 90 kDa

    Exposure time: 40s

  • ChIP - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    ChIP - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    Cal-1 cells (Human plasmacytoid dendritic cell line) were cross-linked with 1% formaldehyde for 5 min at RT. Cross-linked cells were first washed with ice-cold PBS and then resuspended in ice-cold RIPA buffer (10mM Tris-HCl pH8, 140 mM NaCl, 1mM EDTA pH 8, 0.5 mM EGTA, 1% Triton X-100, 0.1% SDS and 0.1% Sodium Deoxycholate) to a final concentration of 5x106 cells/ml. DNA was sheared with a Misonix XL sonicator, by performing 12 x 45'' sonication cycles at power setting of 5. For each ChIP reaction, 2x107 chromatin cell equivalents were incubated overnight with10 μg of ab217668. The following day, chromatin/antibody complexes were incubated with 50 μl of Protein G/Protein A magnetic beads mix (G to A ratio 3:1) for 4 h at 4°C. Normal rabbit IgG was added to the beads as control.

    The TCF-4 locus ChIP-seq tracks for BRD4 (blue), RNA Pol2 (red), and TCF-4 (green) are shown for Cal-1 cells. This data was kindly provided by our collaborator Dr. Louis M. Staudt, and has been published (PMID: 27846392).

  • Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    Blocking and Diluting buffer and concentration: 5% NFDM /TBST

    The isoforms expression pattern is consistent with the literatures (PMID: 21789225).

    All lanes: Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668) at 1/200 dilution

    All lanes: SH-SY5Y (Human neuroblastoma cell line from bone marrow) nuclear extracts at 10 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

    Predicted band size: 71 kDa

    Observed band size: 90 kDa

    Exposure time: 3min

  • Flow Cytometry (Intracellular) - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    Intracellular flow cytometric analysis of 1% paraformaldehyde-fixed, ice-cold methanolpermeabilized Cal-1 cells (Human plasmacytoid dendritic cell line) (black - positive control) and Cal-1 cells infected with either Ctrl (left green) or TCF-4 (right green) shRNA, labeling TCF-4 with ab217668 at 1/100 dilution (green and black) compared with a Rabbit IgG control (grey). Goat anti-Rabbit IgG (Alexa Fluor® 647) at 1/500 dilution was used as the secondary antibody.

    The data was provided by our collaborator Dr. Louis M. Staudt, and published in Cancer Cell 30, 764-778, 2016 (PMID: 27846392). Several TCF4 shRNAs were used. This FC image shows data for shRNA TCF4

    1.

  • Flow Cytometry (Intracellular) - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    Flow cytometry overlay histogram showing SH-SY5Y cells stained with ab217668 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in nan followed by the antibody (ab217668) (1x 106in 100µl at 0.1µg/ml (1/19400)) for 30min at 22°C.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

    Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

    This antibody gave a positive signal in SH-SY5Y Fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

  • Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

    All lanes: Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668) at 1/1000 dilution

    Lane 1: U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate at 20 µg

    Lane 2: Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 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

    Developed using the ECL technique.

    Predicted band size: 71 kDa

    Observed band size: 58 kDa, 79 kDa

    Exposure time: 3min

  • Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668), expandable thumbnail

    Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

    All lanes: Western blot - Anti-TCF-4 antibody [NCI-R159-6] (ab217668) at 1/1000 dilution

    Lane 1: Daudi (human Burkitt's lymphoma lymphoblast) cytoplasmic fraction at 20 µg

    Lane 2: Daudi nuclear fraction at 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

    Developed using the ECL technique.

    Predicted band size: 71 kDa

    Observed band size: 58 kDa, 79 kDa

    Exposure time: 3min

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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