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.
Host cell line: Sf9 cells
Formulation
40 mM Tris-HCl, pH 8.0,110 mM NaCl, 2.2 mM KCl, 200 mM imidazole,and 20% glycerol.
Species
Human
Application
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling
Notes
50 ul reaction mix (20 mM phosphate pH 7.4, 0.05% Tween-20, 40 uM 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.