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.

Limited Uses

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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.