Revision 7

#2569Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IP, ChIP, ChIP-seq, C&R, C&T

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

58, 79

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9NQB0

Entrez-Gene Id:

6934

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:50
CUT&Tag 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

TCF4/TCF7L2 (C48H11) Rabbit mAb detects endogenous levels of total TCF4/TCF7L2 protein.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse, Chicken

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu331 of human TCF4/TCF7L2.

Background

LEF1 and TCF are members of the high mobility group (HMG) DNA-binding protein family of transcription factors that consists of the following: Lymphoid Enhancer Factor 1 (LEF1), T Cell Factor 1 (TCF1/TCF7), TCF3/TCF7L1, and TCF4/TCF7L2 (1). LEF1 and TCF1/TCF7 were originally identified as important factors that regulate early lymphoid development (2) and act downstream in Wnt signaling. LEF1 and TCF bind to Wnt response elements to provide docking sites for β-catenin, which translocates to the nucleus to promote the transcription of target genes upon activation of Wnt signaling (3). LEF1 and TCF are dynamically expressed during development and aberrant activation of the Wnt signaling pathway is involved in many types of cancers, including colon cancer (4,5).

TCF4/TCF7L2 is expressed widely during development. Gene targeting studies indicate that TCF4/TCF7L2 is required to maintain the crypt stem cells of the small intestine (6,7). TCF4/TCF7L2 has several splicing isoforms that are expressed differentially in tissues and during cancer progression (8,9). Studies also indicate that a variant of the TCF4/TCF7L2 gene confers an increased risk of type 2 diabetes (10).

  1. Waterman, M.L. (2004) Cancer Metastasis Rev 23, 41-52.
  2. Schilham, M.W. and Clevers, H. (1998) Semin Immunol 10, 127-32.
  3. Brantjes, H. et al. (2002) Biol Chem 383, 255-61.
  4. Reya, T. and Clevers, H. (2005) Nature 434, 843-50.
  5. Logan, C.Y. and Nusse, R. (2004) Annu Rev Cell Dev Biol 20, 781-810.
  6. Cho, E.A. and Dressler, G.R. (1998) Mech. Dev. 77, 9-18.
  7. Korinek, V. et al. (1998) Nat. Genet. 19, 379-383.
  8. Howng, S.L. et al. (2004) Int. J. Oncol. 25, 1685-1692.
  9. Shiina, H. et al. (2003) Clin. Cancer Res. 9, 2121-2132.
  10. Grant, S.F. et al. (2006) Nat. Genet. 38, 320-323.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN C&T: CUT&Tag

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 7
#2569

TCF4/TCF7L2 (C48H11) Rabbit mAb

Western Blotting Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell types using TCF4/TCF7L2 (C48H11) Rabbit mAb.
Western Blotting Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
Simple Western analysis of lysates (0.1 mg/mL) from HepG2 cells using TCF4/TCF7L2 (C48H11) Rabbit mAb #2569. The virtual lane view (left) shows the target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess ​​​​​​​Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

No image available
Chromatin Immunoprecipitation Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either TCF4/TCF7L2 (C48H11) Rabbit mAb or Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. TCF4/TCF7L2 and β-Catenin are known to associate with each other on chromatin. The figure shows binding of both TCF4/TCF7L2 and β-Catenin across CAMK2D, a known target gene of both TCF4/TCF7L2 and β-Catenin (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
Chromatin Immunoprecipitation Image 2: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either TCF4/TCF7L2 (C48H11) Rabbit mAb or Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. TCF4/TCF7L2 and β-Catenin are known to associate with each other on chromatin. The figure shows binding of both TCF4/TCF7L2 and β-Catenin across chromosome 4 (upper), including CAMK2D (lower), a known target gene of both TCF4/TCF7L2 and β-Catenin (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and either TCF4/TCF7L2 (C48H11) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CaMK2D Intron 3 Primers #5111, human c-Myc promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT and RUN Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
CUT&RUN was performed with HCT 116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across MYC, a known target gene of TCF4 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 2: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
CUT&RUN was performed with HCT 116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 8 (upper), including MYC (lower), a known target gene of TCF4 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 3: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
CUT&RUN was performed with HCT 116 cells and either TCF4/TCF7L2 (C48H11) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human PRKACB intron 1 primers, human MYC exon 1 primers, and human ITM2A upstream primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT and Tag Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
CUT&Tag was performed with HCT 116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across MYC, a known target gene of TCF4 (see our ChIP-qPCR figure).
CUT and Tag Image 2: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
CUT&Tag was performed with HCT116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 8 (upper), including MYC (lower), a known target gene of TCF4 (see our ChIP-qPCR figure).