Products from BPS Bioscience require a minimum order value above 400€Encompassing Amino Acids: 2-end (all components)
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 50 µl reaction mix (20 mM phosphate pH 7.4, 0.05% Tween-20, 40 µM S-adenosylmethionine, and 250-2000 ng enzyme complex) is added to the wells coated with histone H3 substrate on a Neutravidin black plate. 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. Add HRP chemiluminescent substrates and read luminescence.
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.
Description: Complex of human EZH2 (GenBank Accession No. NM_004456), a.a. 2-end with N-terminal His-tag, MW= 86 kDa, human EED (GenBank Accession No. NM_003797), a.a. 2-end with N-terminal FLAG-tag, MW= 51 kDa, and human SUZ12 (GenBank Accession No. NM_015355), a.a. 2-end with N-terminal His-tag, MW = 87 kDa, co-expressed in a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, and 20% Glycerol.
Genbank: NM_004456 (EZH2), NM_003797(EED), NM_015355 (SUZ12)
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal His-tag (EZH2, SUZ12), N-terminal FLAG-tag (EED)
Uniprot: Q15910
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Varambally, S. et al. Science 322 (5908), 1695-1699 (2008).
2. Rakotobe, D. et al., Virol. J. 5, 32 (2008). Application Reference:
Pint lincRNA connects the p53 pathway with epigenetic silencing by the Polycomb repressive complex 2 (2013)
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