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Recombinant Proteins & Enzymes

Recombinant PRC2 complex

Catalog No: 31387 Format: 20 µg $425 Buy Now
Catalog No: 31887 Format: 1 mg $3,800 Buy Now
Expressed In: Baculovirus Protein Species: Human

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for Recombinant PRC2 complexContents

Recombinant PRC2 complex expressed in Sf9 cells and provided in 25 mM HEPES pH 7.5, 300 mM NaCl, 5% Glycerol, 0.04% Triton X-100, 0.2 mM TCEP, 0.2 mg/ml 3x Flag peptide. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant PRC2 complexBackground

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.

for Recombinant PRC2 complexApplication 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.

for Recombinant PRC2 complexProtein 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.

 
PRC2 complex Coomassie gel

Recombinant PRC2 complex gel.
PRC2 complex was run on an 8% SDS-PAGE gel and stained with Coomassie blue.

PRC2 complex activity assay

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.

PRC2 complex HTRF assay

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.

for Recombinant PRC2 complexStorage

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.

for Recombinant PRC2 complexGuarantee

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.

 

Technical Data Sheet

Recombinant PRC2 complex
 

Data Thumbnails

PRC2 complex Coomassie gel

PRC2 complex Coomassie stained gel.
(Click image to enlarge and see details).

PRC2 complex activity assay

PRC2 complex activity assay.
(Click image to enlarge and see details).

PRC2 complex HTRF assay

PRC2 complex HTRF assay.
(Click image to enlarge and see details).