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Key Documents

ABE2945

Sigma-Aldrich

Anti-phospho EZH2 (Thr345)

Sinónimos:

Histone-lysine N-methyltransferase EZH2;EC:2.1.1.356;ENX-1;Enhancer of zeste homolog 2;Lysine N-methyltransferase 6

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.43

biological source

rabbit

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

mol wt

calculated mol wt 85.36 kDa
observed mol wt ~85 kDa

purified by

affinity chromatography

species reactivity

human

species reactivity (predicted by homology)

feline, frog, bovine, canine, mouse, monkey

packaging

antibody small pack of 100 μL

technique(s)

immunocytochemistry: suitable
western blot: suitable

isotype

IgG

epitope sequence

Internal

Protein ID accession no.

UniProt accession no.

storage temp.

2-8°C

target post-translational modification

phosphorylation (pThr345)

Gene Information

human ... EZH2(2146)

General description

Histone-lysine N-methyltransferase EZH2 (UniProt: Q15910; also known as EC:2.1.1.356, ENX-1, Enhancer of zeste homolog 2, Lysine N-methyltransferase 6, EZH2) is encoded by the EZH2 (also known as KMT6) gene (Gene ID: 2146) in human. EZH2 is a polycomb group protein that is expressed in many tissues. It is the catalytic subunit of the PRC2/EED-EZH2 complex that methylates lysine 9 (H3K9me) and lysine 27 (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. It is shown to mono-, di- and trimethylate lysine 27 of histone H3 (H3K27me1, H3K27me2 and H3K27me3). It displays a preference for substrates with less methylation and loses activity when progressively more methyl groups are incorporated into H3K27, H3K27me0 > H3K27me1 > H3K27me2. EZH2 phosphorylation by Akt1 reduces its methyltransferase activity. Its phosphorylation at threonine 345 by CDK1 and CDK2 is shown to promote maintenance of H3K27me3 levels at EZH2-target loci, thus leading to epigenetic gene silencing. Compared to EZH1-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. It is reported to regulate the circadian clock via histone methylation at the promoter of the circadian genes and is essential for the CRY1/2-mediated repression of the transcriptional activation of PER1/2 by the CLOCK-ARNTL/BMAL1 heterodimer. Overexpression of EZH2 has been reported in numerous tumors, including carcinomas of the breast, colon, larynx, testis, and lymphoma. Mutations in EZH2 gene are known to cause Weaver syndrome, a disease characterized by accelerated growth and osseous maturation and unusual craniofacial appearance. (Ref.: Chen, S., et al. (2010). Nat. Cell Biol. 12(11); 1108-1114; Etchegaray, J-P., et al. (2006). J. Biol. Chem. 281(30); 21209-21215; Cha, T-L., et al. (2005). Science. 310(5746); 306-310).

Specificity

This rabbit polyclonal antibody specifically detects Histone-lysine N-methyltransferase EZH2 phosphorylated on threonine 345.

Immunogen

KLH-conjugated linear peptide corresponding to 12 amino acids surrounding phosphothreonine 345 from the internal region of human Histone-lysine N-methyltransferase EZH2.

Application

Quality Control Testing

Evaluated by Western Blotting in LNCaP cell lysate.

Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected EZH2 phosphorylated on threonine 345 in LNCaP cell lysate, but not in lysates treated with protein phosphatase (400 units for 30 min at 30°C).

Tested applications

Immunocytochemistry Analysis: A representative lot detected phospho EZH2 (Thr345) in Immunocytochemistry applications (Chen, S., et. al. (2010). Nat Cell Biol. 12(11):1108-14).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Physical form

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide

Reconstitution

0.1 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


Certificados de análisis (COA)

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