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Encompassing Amino Acids: 2-end (all components)
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 50 µl reaction mix (20 mM phosphate pH 7.4, 0.05% Tween-20, 40 µM S-adenosylmethionine, and enzyme complex) add to the wells coated with the substrate on a Neutravidin white plate (G-Biosciences). Incubate for 1hr. Add antibody against methylated K27 residue of histone H3, incubate 1 hr. Then, add secondary HRP-labeled antibody and incubate 30 min. Finally, add HRP chemiluminescent substrates and read luminescence.
Description: Complex of human EZH1, GenBank Accession No. NM_001991, a.a. 2-end, with N-terminal His-tag, MW= 86 kDa, human EED, NM_003797, a.a. 2-end, with N-terminal FLAG-tag, MW= 51 kDa, human SUZ12, NM_015355, a.a. 2- end, with N-terminal His-tag, MW = 87 kDa, human AEBP2, NM_153207, a.a. 2-end, with N-terminal His-tag, MW= 53 kDa, and human RbAp48, NM_005610, a.a. 2-end, with N-terminal His-tag, MW = 48 kDa, co-expressed in baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0,110 mM NaCl, 2.2 mM KCl, 200 mM imidazole,and 20% glycerol.
Genbank: NM_001991 (EZH1), NM_003797(EED), NM_015355 (SUZ12), NM_153207(AEBP2), NM_005610(RbAP48)
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal His-tag (EZH1, SUZ12, AEBP2, RbAp48), N-terminal FLAG-tag (EED)
Uniprot: Q92800
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Margueron R. et al. Mol. Cell 32, 4, 503-518, 2008.
2. Shen X. et al., Mol. Cell 32, 4, 491-502, 2008.
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