For the
identification of methylations in the recombinant PRC2-PALI1PIR-long
complex, 0.5 µM protein was incubated in the presence or absence of 20 µM SAM
(NEB #B9003) in 25 mM HEPES pH 8.0, 50 mM NaCl, 2 mM MgCl2, 2 mM 2-mercaptoethanol
and 10 % glycerol for 1 hour at 30 °C, in a total final volume of 85 µL.
The proteins were then alkylated, subjected to tryptic digestion and the
tryptic peptides where treated and analysed by MS/MS.