Overview

  • Product name

    Anti-TCF-4 antibody [NCI-R159-6]
    See all TCF-4 primary antibodies
  • Description

    Rabbit monoclonal [NCI-R159-6] to TCF-4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, ChIP, IHCmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human TCF-4 aa 350-550. The exact sequence is proprietary.
    Database link: P15884

  • Positive control

    • WB: Nuclear extracts of Cal-1 cells infected with control TCF-4 shRNA. SH-SY5Y nuclear extracts. IHC: Human tonsil. Blastic plasmacytoid dendritic cell neoplasm (BPDCN) cell pellets after selection and induction of shRNA expression for 1 day. Flow cytometry: Cal-1 cells. ChIP: Cal-1 cells.
  • General notes

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

    A Druggable TCF4- and BRD4-Dependent Transcriptional Network Sustains Malignancy in Blastic Plasmacytoid Dendritic Cell Neoplasm

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab217668 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
WB 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa).
Flow Cyt 1/100.
ChIP Use a concentration of 2.5 µg/ml.
IHC 1/100.

Fixative: 4% Formalin

Antigen retrieval: Low pH retrieval solution

Target

  • Function

    Transcription factor that binds to the immunoglobulin enchancer Mu-E5/KE5-motif. 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'.
  • Tissue specificity

    Expressed in adult heart, brain, placenta, skeletal muscle and to a lesser extent in the lung. In developing embryonic tissues, expression mostly occurs in the brain.
  • Involvement in disease

    Defects in TCF4 are a cause of Pitt-Hopkins syndrome (PTHS) [MIM:610954]. PTHS is a rare syndromic encephalopathy characterized by severe psychomotor delay, epilepsy, daily bouts of diurnal hyperventilation starting in infancy, mild postnatal growth retardation, postnatal microcephaly, and distinctive facial features. Since most hitherto reported cases have been sporadic, with males and females equally affected, PTHS is regarded as an autosomal dominant condition.
  • Sequence similarities

    Contains 1 basic helix-loop-helix (bHLH) domain.
  • Domain

    the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • bHLHb19 antibody
    • Class B basic helix-loop-helix protein 19 antibody
    • E2 2 antibody
    • FECD3 antibody
    • Immunoglobulin transcription factor 2 antibody
    • ITF 2 antibody
    • ITF-2 antibody
    • ITF2 antibody
    • ITF2_HUMAN antibody
    • MGC149723 antibody
    • MGC149724 antibody
    • PTHS antibody
    • SEF 2 antibody
    • SEF-2 antibody
    • SEF2 1 antibody
    • SEF2 1A antibody
    • SEF2 1B antibody
    • SEF2 1D antibody
    • SEF2 antibody
    • SL3 3 enhancer factor 2 antibody
    • SL3-3 enhancer factor 2 antibody
    • TCF 4 antibody
    • TCF-4 antibody
    • TCF4 antibody
    • Transcription factor 4 antibody
    • Transcription factor 4, isoform C antibody
    • Transcription factor 4, isoform D antibody
    • Transcription factor 4, isoform E antibody
    • Transcription factor 4, isoform L antibody
    • Transcription factor 4, isoform M antibody
    • Transcription factor 4, isoform R antibody
    see all

Images

  • All lanes : 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
    Lane 2 : Nuclear extracts of Cal-1 cells infected with TCF4 shRNA

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 71 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?


    Exposure time: 40 seconds


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

  • 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.

  • 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).

  • Flow cytometric analysis of 1% paraformaldehyde-fixed, ice-cold methanol permeabilized 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.

     

  • 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).

     

  • Anti-TCF-4 antibody [NCI-R159-6] (ab217668) at 1/200 dilution + SH-SY5Y (Human neuroblastoma cell line from bone marrow) nuclear extracts at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 71 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


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

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

References

This product has been referenced in:

  • Liang TS  et al. MicroRNA-506 inhibits tumor growth and metastasis in nasopharyngeal carcinoma through the inactivation of the Wnt/ß-catenin signaling pathway by down-regulating LHX2. J Exp Clin Cancer Res 38:97 (2019). Read more (PubMed: 30791932) »
  • Li P  et al. Clinical significance and biological role of Wnt10a in ovarian cancer. Oncol Lett 14:6611-6617 (2017). Read more (PubMed: 29163692) »
See all 4 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab217668.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up