Revision 12

#93212Store 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-IC, ChIP, ChIP-seq

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

180

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q15047

Entrez-Gene Id:

9869

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
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:1000
Immunofluorescence (Immunocytochemistry) 1:800
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

ESET (D4M8R) XP® Rabbit mAb recognizes endogenous levels of total ESET protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human ESET protein.

Background

The Erg-associated protein with SET domain (ESET), also known as SET-domain, bifurcated 1 (SETDB1) protein, is a member of a family of histone lysine methyltransferases, each of which contains a conserved catalytic SET domain originally identified in Drosophila Su[var]3-9, Enhancer of zeste, and Trithorax proteins (1). ESET also contains tudor and methyl-CpG-binding domains, which may coordinate binding to methylated histones and methylated DNA, respectively (1). ESET methylates histone H3 Lys9, creating a transcriptionally repressive mark that facilitates gene silencing (1-3). However, unlike SUV39H histone H3 Lys9 methyltransferases, which function mainly in heterochromatin regions such as pericentric heterochromatin, ESET functions mainly in euchromatic regions to repress gene promoters (3). ESET interacts with a variety of proteins, including transcription factors (ERG), histone deacetylases (HDAC1/2), DNA methyltransferases (DNMT3A/B) and transcriptional co-repressors (mSin3A/B, MBD1, KAP-1, the ATFa-associated modulator mAM) (1-6). mAM forms a complex with ESET, stimulating its methyltransferase activity, specifically the conversion of di-methyl to tri-methyl histone H3 Lys9 (2). MBD1 recruits ESET to the CAF-1 complex to facilitate methylation of histone H3 Lys9 during replication-coupled chromatin assembly in S phase (5). DNMT3A recruits ESET to silenced promoters in cancer cells (7). ESET may play a role in the pathogenesis of Huntington's disease, since levels of ESET protein and tri-methyl histone H3 Lys9 are both increased in diseased brains (8).

  1. Yang, L. et al. (2002) Oncogene 21, 148-152.
  2. Wang, H. et al. (2003) Mol. Cell 12, 475-487.
  3. Schultz, D.C. et al. (2002) Genes Dev. 16, 919-932.
  4. Yang, L. et al. (2003) Biochem. J. 369, 651-657.
  5. Sarraf, S.A. and Stancheva, I. (2004) Mol. Cell 15, 595-605.
  6. Ichimura, T. et al. (2005) J. Biol. Chem. 280, 13928-13935.
  7. Li, H. et al. (2006) J. Biol. Chem. 281, 19489-19500.
  8. Ryu, H. et al. (2006) Proc. Natl. Acad. Sci. USA 103, 19176-19181.

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-IC: Immunofluorescence (Immunocytochemistry) 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.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
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 12
#93212

ESET (D4M8R) XP® Rabbit mAb

Western Blotting Image 1: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 ESET (D4M8R) XP® Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
No image available
Immunohistochemistry Image 1: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 ESET (D4M8R) XP® Rabbit mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析。
Immunohistochemistry Image 2: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 ESET (D4M8R) XP® Rabbit mAb 对石蜡包埋的人非小细胞肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 ESET (D4M8R) XP® Rabbit mAb 对石蜡包埋的人非小细胞肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 ESET (D4M8R) XP® Rabbit mAb,对石蜡包埋的人类非霍奇金淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 5: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 ESET (D4M8R) XP® Rabbit mAb 对石蜡包埋的人前列腺癌细胞进行免疫组织化学分析。
Immunofluorescence Image 1: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 ESET (D4M8R) XP® Rabbit mAb(绿色)对 MCF7 细胞进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)进行标记。
Chromatin Immunoprecipitation Image 1: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 K-562 细胞中提取的交联染色质和 ESET (D4M8R) XP® Rabbit mAb 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 ZNF12 基因内结合作用。如需了解其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 4 x 106 K-562 细胞中提取的交联染色质和 ESET (D4M8R) XP® Rabbit mAb 进行染色质免疫沉淀分享。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 7(上图)内的结合,包括 ZNF12 基因(下图)。
Chromatin Immunoprecipitation Image 3: ESET (D4M8R) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 K-562 细胞中提取的交联染色质,在加入 ESET (D4M8R) XP® Rabbit mAb 或 Normal Rabbit IgG #2729 后,进行染色质免疫沉淀分析。使用 SimpleChIP® Human GAPDH Exon 1 Promoter Primers #5516、SimpleChIP® Human β-Actin Promoter Primers #13653 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。