EZH2 (Y641F) /EED/SUZ12/RbAp48/AEBP2, His-tag, FLAG-tag Recombinant

Catalog #

51017

$385 *

Size: 20 µg

Qty

Bulk/Custom

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Description

Mutant version of our regular EZH2 5-member complex (Cat. #51004), but with a Tyr-to-Phe mutation on a.a. 646 of the EZH2 protein. Complex of human EZH2 (GenBank Accession No. NM_004456), a.a. 2-end; Y646F*, with N-terminal His tag, MW= 86 kDa, human EED (NM_003797), a.a. 2-end with N-terminal Flag tag, MW= 51 kDa, human SUZ12 (NM_015355), a.a. 2-end with N-terminal His tag, MW = 87 kDa, Human AEBP2 (NM_153207), a.a. 2-end with N-terminal His tag, MW= 53 kDa, and human RbAp48 (NM_005610), a.a. 2-end with N-terminal His tag, MW = 48 kDa, co-expressed in baculovirus expression system.

This product has been cited 1 time.

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Synonyms

KTM6A, EZH2, PRC2, Enhancer Of Zeste Homolog 2, EZH2 (Y646F), EZH , Retinoblastoma Binding Protein 4 (RBBP4)

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Product Info

Storage and Usage

Citations1

Species

Human

Host Species/Expression System

Sf9 insect cells

Format

Aqueous buffer solution

Formulation

25 mM Tris-HCl, pH 8.0, 138 mM NaCl, 0.05% Tween-20, and 10% glycerol.

325 kDa (complex)

Amino Acids

2–end (all components)

Genbank #

NM_004456 (EZH2), NM_003797(EED), NM_015355 (SUZ12), NM_153207(AEBP2), NM_005610(RbAP48)

UniProt #

Q15910

Tag(s)

N-terminal His-tag (EZH2, SUZ12, RbAp48, AEBP2), N-terminal FLAG-tag (EED)

Background

EZH2 is the catalytic subunit of the PRC2/EED-EZH2 complex, which methylates Lys9 (H3K9me) and Lys27 (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate Lys27 of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-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. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4.

References

1. Varambally, S. et al. Science 322 (5908), 1695-1699 (2008).
2. Rakotobe, D. et al., Virol. J. 5, 32 (2008).

Application Reference(s):
1. Arcand, M., et al., "Development of High-Throughput Assays to Study Methylases, Demethylases and Deacetylases." Poster presented as part of the BioTechniques Fall 2011 Epigenetics Meeting.
2. Yap, D.B, et al., "Somatic mutations at EZH2 Y641 act dominantly through a mechanism of selectively altered PRC2 catalytic activity, to increase H3K27 trimethlyation." (2010). Blood. 117: 2451-2459.

Storage/Stability

≥ 6 months at -80°C

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for Western blot analysis and to investigate protein-protein interactions in the EZH2 complex.

Shipping Temperature

-80°C

Notes

This mutation is often cited in the scientific literature as Y641F in variant 3 (GenBank #NM_001203247).

Warnings

Avoid freeze/thaw cycles

Scientific Category

Methyltransferase

Regulatory Status

Research Use Only

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Datasheets/Protocols

51017_Lot120227.pdf 51017_Lot120808.pdf

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EZH2 (Y641F) /EED/SUZ12/RbAp48/AEBP2, His-tag, FLAG-tag Recombinant

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