Amplify namp100v solution

The peptide arrays were pre-incubated for 10 min in methylation buffer (20 mM HEPES pH8, 50 mM NaCl, 5 mM DTT) and then incubated in the methylation buffer containing 300 nM SET8 and 0.76 μM labeled [methyl-³H]-AdoMet for 60 min. The membranes were washed 5 times for 5 minutes with wash buffer (100 mM NH₄HCO₃ and 1% SDS). Finally, the peptide arrays were incubated with Amplify NAMP100V solution (GE Healthcare, Munich, Germany) for 5 minutes. The peptide arrays were exposed to Hyperfilm TM high performance autoradiography film (GE Healthcare, Munich, Germany) in the dark at −80 °C for 7–10 days. The autoradiography films were developed in Optimus TR developing machine.

Peptide SPOT arrays were preincubated in methylation buffer containing 20 mM Tris–HCl (pH 9) and 5 mM DTT for 5 min on a shaker. Afterward, the SPOT arrays were incubated in methylation buffer containing additionally 50 nM SETD6 and 0.76 μM labeled [methyl-³H]-AdoMet (PerkinElmer, Inc) for 1 h at room temperature. Next, the arrays were washed five times for 5 min with 100 mM NH₄HCO₃ and 1% SDS. After washing, the arrays were incubated in Amplify NAMP100V solution (GE Healthcare) for 5 min. Then, the arrays were exposed to Hyperfilm high-performance autoradiography films (GE Healthcare) in the dark at −80 °C for different exposure times and developed using an Optimus TR developing machine.