Recombinant Anti-TCF-4 antibody [NCI-R159-6] (ab217668)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [NCI-R159-6] to TCF-4
- Suitable for: Flow Cyt (Intra), WB, ChIP, IHC
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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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 -
Specificity
Mouse specificity in WB only.
Weak human specificity in IHC.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, ChIP, IHCmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Nuclear extracts of Cal-1 cells infected with control TCF-4 shRNA. SH-SY5Y nuclear extracts. U-87 MG and Neuro-2a whole cell lysates, Daudi nuclear fraction, U-87 MG whole cell lysate, Neuro-2a whole cell lysate IHC: Human tonsil. Blastic plasmacytoid dendritic cell neoplasm (BPDCN) cell pellets after selection and induction of shRNA expression for 1 day. Flow Cyt (intra): Cal-1 cells. ChIP: Cal-1 cells.
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General notes
For detailed protocol using this antibody for IHC, ChIP, and Flow Cyt, please refer to the following paper:
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
NCI-R159-6 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
1/100.
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WB |
1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa).
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ChIP |
Use a concentration of 2.5 µg/ml.
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IHC |
1/100.
Fixative: 4% FormalinAntigen retrieval: Low pH retrieval solution |
Notes |
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Flow Cyt (Intra)
1/100. |
WB
1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa). |
ChIP
Use a concentration of 2.5 µg/ml. |
IHC
1/100. Fixative: 4% FormalinAntigen retrieval: Low pH retrieval solution |
Target
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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
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Database links
- Entrez Gene: 6925 Human
- Entrez Gene: 21413 Mouse
- Omim: 602272 Human
- SwissProt: P15884 Human
- SwissProt: Q60722 Mouse
- Unigene: 605153 Human
- Unigene: 4269 Mouse
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Alternative names
- bHLHb19 antibody
- Class B basic helix-loop-helix protein 19 antibody
- E2 2 antibody
see all
Images
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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 secondsThe data was provided by the collaborator Dr. Louis M. Staudt, NCI, NIH.
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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.
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All lanes : Anti-TCF-4 antibody [NCI-R159-6] (ab217668) at 1/1000 dilution
Lane 1 : Daudi (human Burkitt's lymphoma lymphoblast) cytoplasmic fraction
Lane 2 : Daudi nuclear fraction
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 58,79 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesExposure time : 3 minutes
Blocking/Dilution buffer: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate.
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All lanes : 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
Lane 2 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 58,79 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesExposure time : 3 minutes
Blocking/Dilution buffer: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate.
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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).
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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.
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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).
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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 minutesBlocking and Diluting buffer and concentration: 5% NFDM /TBST
The isoforms expression pattern is consistent with the literatures (PMID: 21789225).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (25)
ab217668 has been referenced in 25 publications.
- Gong N et al. S100A4/TCF Complex Transcription Regulation Drives Epithelial-Mesenchymal Transition in Chronic Sinusitis Through Wnt/GSK-3β/β-Catenin Signaling. Front Immunol 13:835888 (2022). PubMed: 35154161
- Papes F et al. Transcription Factor 4 loss-of-function is associated with deficits in progenitor proliferation and cortical neuron content. Nat Commun 13:2387 (2022). PubMed: 35501322
- Chan HYE & Chen ZS Multifaceted investigation underlies diverse mechanisms contributing to the downregulation of Hedgehog pathway-associated genes INTU and IFT88 in lung adenocarcinoma and uterine corpus endometrial carcinoma. Aging (Albany NY) 14:7794-7823 (2022). PubMed: 36084949
- Asashima H et al. Impaired TIGIT expression on B cells drives circulating follicular helper T cell expansion in multiple sclerosis. J Clin Invest 132:N/A (2022). PubMed: 36250467
- Jiang L et al. Ivermectin inhibits tumor metastasis by regulating the Wnt/β-catenin/integrin β1/FAK signaling pathway. Am J Cancer Res 12:4502-4519 (2022). PubMed: 36381328