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Recombinant EZH2 protein complex

Expressed In

Baculovirus

Protein Species

Human

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Cat No : 31337

Validation Data Gallery

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

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

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

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

Product Information

Expressed In Baculovirus
Protein Species Human

Contents

25 ug recombinant EZH2 protein complex (contains EZH2, EED and Suz12 proteins) supplied in PBS (10 mM Na2HPO4/KH2PO4), pH 7.4, 2.7 mM kCl, and 138 mM NaCl.

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).

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.