EZH2/EED/SUZ12/RbAp48/AEBP2, FLAG-tag, His-tag

Catalog #: 51004
Size: 50 µg
$775
Species: Human
Host Species: Sf9 insect cells
MW: 325 kDa (complex)
Genbank #: NM_004456 (EZH2), NM_003797(EED), NM_015355 (SUZ12), NM_153207(AEBP2), NM_005610(RbAP48)
Tag(s): N-terminal FLAG-tag(EED), N-terminal His-tag (other components)
a.a: 2–end (all components)
Background:
EZH2 is the catalytic subunit of the PRC2/EED-EZH2 protein 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 protein 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.
Description:
Complex of human EZH2 (GenBank Accession No. NM_004456), a.a. 2-end, 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 a baculovirus expression system.
UniProt
Q15910
Synonym(s):
EZH2 Protein Complex, EZH2, PRC2, Enhancer Of Zeste Homolog 2; RBAP48, Retinoblastoma Binding Protein 4 (RBBP4), NURF55, Chromatin Assembly Factor 1 Subunit C, CAF1 p48; Embryonic Ectoderm Development, hEED, EED, WAIT-1; SUZ12, Suppresso
Specific Activity:
0.08 pmol/min/µg
Assay Conditions:
50 µl reaction mix (20 mM phosphate buffer pH 7.4, 0.05% Tween-20, 20 µM S-adenosylmethionine, and 50-200 ng enzyme complex) add to the wells coated with the substrate. Incubate for 1 hr. Add antibody against methylated K27 residue of histone H3, incubate 1 hr. Then, add secondary HRP-labeled antibody and incubate 30 min. Finally, add HRP chemiluminescent substrates and read luminesence.
Formulation:
40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 200 mM imidazole, and 20% glycerol.
Format:
Aqueous buffer solution
Storage / Stability:

>6 months at -80°C.

Application(s):
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Reference(s):
1. Morin, R., et al. (2010). et al., Nature Genetics. 42, 181 - 185.
2.
Varambally, S. et al. (2008). Science 322 (5908), 1695-1699.
3.
Rakotobe, D. et al. (2008). Virol. J. 5, 32.

Application Reference(s):
1. Arcand, M. et al., Poster presented as part of the BioTechniques Fall 2011 Epigenetics Meeting. , 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. Cohen, A. et al., Weaver Syndrome-Associated EZH2 Protein Variants Show Impaired Histone Methyltransferase Function In Vitro (2016)
3. 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. 

Warning(s):
Avoid freeze/thaw cycles
Scientific Category:
Methyltransferase
Product Type:
Recombinant Protein
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Product Datasheets

51004 Lot 161117.pdf