Cette EZH2 protéine est marqué à la DYKDDDDK Tag,His tag.
Application
Enzyme Activity Assay (EAA)
Attributs du produit
The EZH2 protein complex, active contains EZH2, EED and Suz12 which are the minimal components required for histone methyltransferase activity. EZH2 is specific for H3 lysine 27, a mark of repressed regions of chromatin. 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).
EZH2
Origine: Humain
Hôte: Escherichia coli (E. coli)
Recombinant
> 95 %
WB, SDS, Imm, PC
Indications d'application
Recombinant EZH2 protein complex is suitable for use in histone methyltransferase assays. This protein is useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling. 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.
Restrictions
For Research Use only
Conseil sur la manipulation
Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
Stock
-80 °C
Stockage commentaire
Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation.
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.