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Recombinant PRC2 complex
| Catalog No: 31387 | Format: 20 µg | $510 | Buy |
| Catalog No: 31887 | Format: 1 mg | $4,300 | Buy |
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Expressed In: Baculovirus Protein Species: Human
Contents
A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.
Background
PRC2 (Polycomb Repressive Complex 2) is one of the two classes Polycomb-group, or PcG proteins (the other being PRC1) that are important epigenetic determinants of stem cell identity. They play an important role in long-term epigenetic silencing of genes during cell fate determination and differentiation. PRC2 functions as a repressor of chromatin. PRC2 is required to target recruitment to specific DNA sequences (termed Polycomb Response Elements or PREs) of genomic regions to be silenced. Once associated with chromatin, the PRC2 subunit EZH2 has histone methyltransferase activity that catalyzes the trimethylation of histone H3 at lysine 27. H3K27me3 is well established as a hallmark of regions of repressed chromatin. Trimethylation of lysine 27 leads to the recruitment of PRC1 through the binding of H3K27me3 by chromodomain-containing proteins in PRC1. PRC1 is responsible for long-term gene silencing after cellular differentiation.
Application Notes
PRC2 Complex is suitable for use in the study of enzyme kinetics, inhibitor screening and selectivity profiling.
Catalytic Ability: 17 turnover / enzyme molecule
HMT Assay Conditions: 3.3 μM H3K27me0 peptide was incubated with different concentration of PRC2 Complex in reaction buffer including 50 mM TrisCl, pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 100 μM SAM for 3 hours at room temperature. Activity was detected by HTRF and MALDI-TOF.
Protein Details
Recombinant PRC2 complex that includes full length EZH2, SUZ12, EED and RbAp46/48 (accession numbers NP_001190176.1, NP_056170, NP_003788.2, NP_002884.1, and NP_005601.1, respectively) was expressed in Sf9 and contains an N-terminal FLAG-Tag at the N-terminus of EZH2. The molecular weights of expressed EZH2, SUZ12, EED and RbAp46/48 are 87 kDa, 83 kDa, 50.2 kDa, 47.8 kDa and 47.7 kDa, respectively. The recombinant protein is >95% pure by SDS-PAGE.
References
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Recombinant PRC2 complex gel.
PRC2 complex was run on an 8% SDS-PAGE gel and stained with Coomassie blue.
Recombinant PRC2 complex activity assay.
3.3 μM H3K27me0 peptide was incubates with 40 nM PRC2 complex in reaction buffer including 50 mM TrisCl, pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 100 μM SAM for 3 hours at room temperature.
Recombinant PRC2 complex activity assay.
3.3 μM H3K27me0 peptide was incubated with different concentration of PRC2 complex in reaction buffer for 3 hour at room temperature. PRC2 complex was used in a HTRF assay to determine enzyme linearity. Methylated peptide (H3K27me1) was measured using H3K27me1-specific antibody.
Storage
Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
