Recombinant EZH2 protein complex

Aliases: WVS; ENX1; EZH1; KMT6; WVS2; ENX-1; EZH2b; KMT6A
Catalog No: 31337 Format: 25 µg $485 Buy
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

EZH2, EED and Suz12 are present in the PRC protein complexes that function as mediators of epigenetic transcriptional silencing. EZH2 – (Enhancer of Zeste homolog 2, also designated Enx1 and SETDB1) is a human homolog of the Drosophila Enhancer of Zeste protein. It contains a SET domain that catalyzes the mono-, di- and trimethylation of Histone H3 Lys27. Polycomb group proteins repress gene expression by binding to chromatin and locally altering chromatin structure. Deregulation of EZH2 is linked to a variety of cancers.

Application Notes

Recombinant EZH2 protein complex is suitable for use in histone methyltransferase assays.

Specific Activity: 0.08 pmol/min/μg.

Assay conditions: Prepare a 50 μl reaction mix (20 mM phosphate pH 7.4, 0.05% Tween-20, 40 μM S-adenosylmethionine, and 250-2000 ng enzyme complex) and add to substrate-coated wells with the substrate on a Thermo Neutravidin black plate. Incubate for 1 hr. Add anti-methyl histone H3K27 antibody and incubate 1hr. Next, incubate 1h with the HRP-labeled secondary antibody followed by chemiluminescent detection.

Protein Details

Recombinant human EZH2 (accession number NM_004456, amino acids 2-end with an N-terminal His-Tag, molecular weight 86 kDa) was coexpressed in a baculovirus expression system along with human EED (accession number NM_003797, amino acids 2-end with an N-terminal FLAG-Tag, molecular weight 51 kDa) and human Suz12 (accession number NM_015355, amino acids 2-end with an N-terminal His-Tag, molecular weight 87 kDa).

References

This product was used in the following publications:
Mol. Cell. (2016). 61(2):247-59. PMID: 26778125. (Histone Methyltransferase / HMT Assay)
MAbs. (2018) May/Jun;10(4):539-546. PMID: 29485921.

 
EZH2 Coomassie gel

EZH2 protein gel.
EZH2 run on an SDS-PAGE gel and stained with Coomassie blue. Arrows indicate recombinant proteins as labeled.

EZH2 activity assay

EZH2 complex activity assay.
Recombinant EZH2 activity measured using a fluorescent histone H3 lysine 27 methyltransferase assay.

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.