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.

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

Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。
Revision 7
#2569

TCF4/TCF7L2 (C48H11) Rabbit mAb

Western Blotting Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
使用 TCF4/TCF7L2 (C48H11) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
Western Blotting Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
使用 TCF4/TCF7L2 (C48H11) Rabbit mAb #2569 对 HepG2 细胞的裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像（左图）显示一抗稀释比例在 1:10 和 1:50 时的靶标条带（如图所示）。对应的电泳图（右图）为一抗稀释比例在 1:10（蓝线）和 1:50（绿线）时沿毛细血管内分子量的化学发光结果。在还原条件下，使用 12-230 kDa 分离模块在 ProteinSimple（BioTechne 品牌）的 Jess™ Simple Western 仪器上进行该实验。

No image available
Chromatin Immunoprecipitation Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，对 HCT116 细胞中提取的交联染色质，在加入 TCF4/TCF7L2 (C48H11) Rabbit mAb 或 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb#8814 后进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。已知 TCF4/TCF7L2 和 β-链蛋白可在染色质上相互结合。图形显示 TCF4/TCF7L2 和 β-链蛋白在 CAMK2D 内结合，已知 CAMK2D 是 TCF4 和 β-链蛋白的一种靶标基因（参见包含 ChIP-qPCR 数据的其他图形）。欲知其他 ChIP-seq 情况，请下载产品说明书。
Chromatin Immunoprecipitation Image 2: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，对 HCT116 细胞中提取的交联染色质，在加入 TCF4/TCF7L2 (C48H11) Rabbit mAb 或 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb#8814 后进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。已知 TCF4/TCF7L2 和 β-链蛋白可在染色质上相互结合。结果图显示 TCF4 和 β-链蛋白在染色体 4（上图）内的结合，包括 TCF4/TCF7L2 和 β-链蛋白的已知靶基因 CAMK2D（下图）（参见包含 ChIP-qPCR 数据的其他结果图）。
Chromatin Immunoprecipitation Image 3: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003对 HCT 116 细胞中提取的交联染色质，在加入 TCF4/TCF7L2 (C48H11) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® Human CaMK2D Intron 3Primers #5111、Human c-Myc promoter primers 与 SimpleChIP® Human α Satellite Repeat Primers #4486，通过实时 PCR 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。
CUT and RUN Image 1: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit # 86652通过 HCT116 细胞和 TCF4/TCF7L2 (C48H11) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。图形显示在 MYC 内结合，MYC 是一种已知的 TCF4 靶标基因（参见包含 CUT&RUN-qPCR 数据的其他图形）。
CUT and RUN Image 2: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit # 86652通过 HCT116 细胞和 TCF4/TCF7L2 (C48H11) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在染色体 8（上图）内的结合，包括 TCF4 的已知靶基因 MYC（下图）（参见包含 CUT&RUN-qPCR 数据的其他结果图）。
CUT and RUN Image 3: TCF4/TCF7L2 (C48H11) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652，通过 HCT 116 细胞和 TCF4/TCF7L2 (C48H11) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN。使用人 PRKACB 内含子 1 引物、人 MYC 外显子 1 引物和人 ITM2A 上游引物通过实时 PCR 对富集的 DNA 进行定量。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。
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).
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Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。