Revision 3

#76010Store 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, FC-FP, ChIP, ChIP-seq

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

25-58

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9UJU2

Entrez-Gene Id:

51176

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.

Application Dilution
Western Blotting 1:1000
Flow Cytometry (Fixed/Permeabilized) 1:100
Chromatin IP 1:50
Chromatin IP-seq 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

LEF1 (D6J2W) Rabbit mAb recognizes endogenous levels of total LEF1 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu80 of human LEF1 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).

  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.

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 FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

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.
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 3
#76010

LEF1 (D6J2W) Rabbit mAb

Western Blotting Image 1: LEF1 (D6J2W) Rabbit mAb Expand Image
使用 LEF1 (D6J2W) Rabbit mAb(上)和 β-Actin (D6A8) Rabbit mAb #8457(下),对不同细胞系的提取物进行蛋白质印迹分析。
Flow Cytometry Image 1: LEF1 (D6J2W) Rabbit mAb Expand Image

以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)作为对照,使用 LEF1 (D6J2W) Rabbit mAb(右图)和共染色的 CD3 (UCHT1) Mouse mAb (APC Conjugate) #19881 对人外周血单核细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。

Chromatin Immunoprecipitation Image 1: LEF1 (D6J2W) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 HCT-116 细胞中提取的交联染色质,在加入 LEF1 (D6J2W) Rabbit mAb 后进行染色质免疫沉淀。使用 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795 制成 DNA 库。结果图显示在 AXIN2 内结合,AXIN2 是一种已知的 LEF1 靶标基因(参见包含 ChIP-qPCR 数据的其他结果图)。如需了解其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: LEF1 (D6J2W) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 HCT-116 细胞中提取的交联染色质,在加入 LEF1 (D6J2W) Rabbit mAb 后进行染色质免疫沉淀。使用 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795 制成 DNA 库。结果图显示在染色体 17(上图)内的结合,包括 LEF1的已知靶基因 AXIN2(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: LEF1 (D6J2W) 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 LEF1 (D6J2W) Rabbit mAb 或 Normal Rabbit IgG #2729 后进行染色质免疫沉淀。使用 SimpleChIP® Human Axin2 Intron 1 Primers #8973 和 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。