WB, IP, ChIP, ChIP-seq, C&R
H M R Mk
Endogenous
98
Rabbit IgG
#Q15910
2146
Product Information
Product Usage Information
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
| CUT&RUN | 1:50 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr345 of human Ezh2 protein.
Background
The polycomb group (PcG) proteins are involved in maintaining the silenced state of several developmentally regulated genes and contribute to the maintenance of cell identity, cell cycle regulation, and oncogenesis (1,2). Enhancer of zeste homolog 2 (Ezh2), a member of this large protein family, contains four conserved regions including domain I, domain II, and a cysteine-rich amino acid stretch that precedes the carboxy-terminal SET domain (3). The SET domain has been linked with histone methyltransferase (HMTase) activity. Moreover, mammalian Ezh2 is a member of a histone deacetylase complex that functions in gene silencing, acting at the level of chromatin structure (4). Ezh2 complexes methylate histone H3 at Lys9 and 27 in vitro, which is thought to be involved in targeting transcriptional regulators to specific loci (5). Ezh2 is deregulated in various tumor types, and its role, both as a primary effector and as a mediator of tumorigenesis, has become a subject of increased interest (6).
Ezh2 is phosphorylated on Thr345 by cyclin-dependent kinase 1 (CDK1), which increases targeting of Ezh2 to its target loci and promotes methylation of H3 at Lys27 (7,8). This activated form of Ezh2 may deposit initial H3K27me3 marks onto newly synthesized daughter strands of DNA during replication (9). In addition, phosphorylation of Thr345 affects Ezh2 stability by promoting its ubiquitination and subsequent degradation (9).
- Sellers, W.R. and Loda, M. (2002) Cancer Cell 2, 349-50.
- Visser, H.P. et al. (2001) Br J Haematol 112, 950-8.
- Chen, H. et al. (1996) Genomics 38, 30-7.
- Tonini, T. et al. (2004) Oncogene 23, 4930-7.
- Müller, J. et al. (2002) Cell 111, 197-208.
- Kleer, C.G. et al. (2003) Proc Natl Acad Sci U S A 100, 11606-11.
- Chen, S. et al. (2010) Nat Cell Biol 12, 1108-14.
- Kaneko, S. et al. (2010) Genes Dev 24, 2615-20.
- Wu, S.C. and Zhang, Y. (2011) J Biol Chem 286, 28511-9.
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 ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN
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
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