Revision 6

#2203Store 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, IP, IHC-P, IF-F, IF-IC, FC-FP, ChIP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

48, 50

Source/Isotype:

Rabbit IgG

UniProt ID:

#P36402

Entrez-Gene Id:

6932

Product Information

Product Usage Information

For a carrier free (BSA and azide free) version of this product see product #85942
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Frozen) 1:800
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200
Chromatin IP 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.

For a carrier free (BSA and azide free) version of this product see product #85942

Specificity / Sensitivity

TCF1/TCF7 (C63D9) Rabbit mAb detects endogenous levels of total TCF1/TCF7 protein. This antibody does not recognize the dominant negative isoforms of TCF1/TCF7 lacking the amino-terminal β-catenin binding domain and does not cross-react with LEF1. TCF1/TCF7 (C63D9) non-specifically labels glomeruli of kidney and fiber-like structures in adipose tissue, skeletal muscle, lung, ovary, colon, and spleen by immunofluorescence.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to a region surrounding Pro96 of human TCF1/TCF7 protein.

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).

TCF1/TCF7 has several isoforms due to alternative splicing and transcription from an alternative promoter. The isoforms generated by the alternative promoter do not contain the amino-terminal β-catenin binding domain and therefore may function in a dominant negative manner (6). TCF1/TCF7 displays dynamic expression both in the total amount and the type of isoforms expressed in T cells during development and differentiation (7).

  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. Waterman, M.L. (2004) Cancer Metastasis Rev. 23, 41-52.
  7. Willinger, T. et al. (2006) J. Immunol. 176, 1439-1446.

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 IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP

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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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 6
#2203

TCF1/TCF7 (C63D9) Rabbit mAb

Western Blotting Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
Western blot analysis of total cell lysates from HT29, Colo201, Jurkat and mouse thymocytes using TCF1/TCF7 (C63D9) Rabbit mAb.
No image available
Immunohistochemistry Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin lymphoma using TCF1/TCF7 (C63D9) Rabbit mAb.
Immunohistochemistry Image 2: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using TCF1/TCF7 (C63D9) Rabbit mAb in the presence of control peptide (left) or TCF1/TCF7 blocking peptide #1007 (right).
Immunohistochemistry Image 3: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human tonsil using TCF1/TCF7 (C63D9) Rabbit mAb.
Immunofluorescence Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen lymph node from wild-type (left) Tcf7GFP flox (right; Jax Strain 030909) mice using TCF1/TCF7 (C63D9) Rabbit mAb #2203 (red) and CD4 (RM4-5) Rat mAb (redFluor 710 Conjugate) #75508 (blue). EGFP insertion around Tcf7 exon 2 interferes with expression of long isoforms, but not short isoforms.
Immunofluorescence Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of DLD-1 cells using TCF1/TCF7 (C63D9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
Flow Cytometry Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
Flow cytometric analysis of A20 cells (blue) and EL-4 cells (green) using TCF1/TCF7 (C63D9) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Chromatin Immunoprecipitation Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from mouse thymocytes and either TCF1/TCF7 (C63D9) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse LEF1 Upstream Primers #80993 and SimpleChIP® Mouse MYT-1 Promoter Primers #8985. 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.