EZH2 Assay Service
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Target
EZH2
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Description
Screening and/or profiling inhibitor compounds against EZH2 methyltransferase activity in a biochemical assay.
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Synonyms
KTM6A, EZH2, PRC2, Enhancer Of Zeste Homolog 2, EZH
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Example Data
*Example only, final data may vary.
Assay Details
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Assay Format
Chemiluminescent
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Reference Compounds and IC50
GSK343, 10 nM
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Assay Principle
The EZH2 Chemiluminescence Assay is an EZH2/EED/SUZ12/RbAp48/AEBP2 chemiluminescence three step kit designed to measure activity of the EZH2 (enhancer of zeste homolog 2) complex (EZH2/EED/SUZ12/RbAp48/ AEBP) for screening and profiling purposes. The EZH2 Assay is performed in a 96-well plate. The EZH2 Activity Assay uses a highly specific antibody that recognizes methylated Histone H3 on K27. In the first step, S-adenosylmethionine is incubated with methyltransferase and sample prepared in assay buffer for one hour. This is followed by addition of the primary antibody. Finally, the plate is treated with an HRP-labeled secondary antibody and addition of the HRP substrate to produce chemiluminescence signal proportional to the EZH2 activity.
Target Details
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Protein Family
Methyltransferases
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UniProt
Q15910
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Background
EZH2 (enhancer of zeste homolog 2) is a histone-lysine N-methyltransferase enzyme, that acts by adding methyl groups to the lysine 27 (K27) of histone H3, making it a silent chromatin. It is a functional unit of the larger complex PRC2 (polycomb repressive complex 2), which also includes EED, Suz12, AEBP2 (adipocyte enhancer-binding protein) and RbAp48 (histone-binding protein RBBP4). PRC2 is crucial for epigenetic regulation and is involved in stem cell differentiation and embryonic development. Abnormalities in PRC2 result in cancer since this complex also promotes double strand DNA repair. EZH2 is seen as an attractive target in cancer therapy, as its levels are elevated in multiple cancer types (example, breast, renal cancer, melanoma, and lymphoma). The development of inhibitors for EZH2 and PRC2 is a promising area of research for the treatment of cancer.
Delivery
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Estimated Turnaround
Two to three weeks following delivery of compounds
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Results
Extensive report with raw and analyzed data, graphs, and detailed protocols. Includes positive control for inhibition.
