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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 6
#2203

TCF1/TCF7 (C63D9) Rabbit mAb

Western Blotting Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
使用 TCF1/TCF7 (C63D9) Rabbit mAb,对来自 HT29、Colo201、Jurkat 和小鼠胸腺细胞的总细胞裂解物进行蛋白质印迹分析。
No image available
Immunohistochemistry Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
使用 TCF1/TCF7 (C63D9) Rabbit mAb 对石蜡包埋的人非霍奇金淋巴瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 2: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
在对照肽(左图)或 TCF1/TCF7 Blocking Peptide #1007(右图)存在的情况下,使用 TCF1/TCF7 (C63D9) Rabbit mAb 对石蜡包埋的人肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 3: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
使用 TCF1/TCF7 (C63D9) Rabbit mAb 对石蜡包埋的人扁桃体组织进行免疫组织化学分析。
Immunofluorescence Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
使用 TCF1/TCF7 (C63D9) Rabbit mAb #2203红色)和 CD4 (RM4-5) Rat mAb (redFluor 710 Conjugate) #75508(蓝色),对野生型(左图)Tcf7GFP flox(右图;Jax 品系 030909)小鼠的固定冷冻淋巴结进行共聚焦免疫荧光分析。在 Tcf7 外显子 2 周围插入 EGFP 干扰了长同工型表达,但不干扰短同工型表达。
Immunofluorescence Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
使用 TCF1/TCF7 (C63D9) Rabbit mAb(绿色)对 DLD-1 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 Alexa Fluor®555 phalloidin(红色)进行标记。
Flow Cytometry Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
使用 TCF1/TCF7 (C63D9) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 A20 细胞(蓝色)和 EL-4 细胞(绿色)进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
Chromatin Immunoprecipitation Image 1: TCF1/TCF7 (C63D9) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对小鼠胸腺细胞的交联染色质和 TCF1/TCF7 (C63D9) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀分析。使用 SimpleChIP® Mouse LEF1 Upstream Primers #80993 和 SimpleChIP® Mouse MYT-1 Promoter Primers #8985 进行实时 PCR 来对富集的 DNA 进行定量分析。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。