Lysine Methylation

From CCP4 wiki
  • 1mg/ml protein
  • 50mM HEPES (pH7.5)
  • 250mM NaCl
  • Do NOT use Tris

Add per 1ml protein:

  • 20μl 1M dimethylamine-borane complex (ABC) – freshly prepared
  • 40μl 1M formaldehyde (from 37% stock)

Mix gently and incubate for 2hrs at 4°C

Add again:

  • 20μl 1M dimethylamine-borane complex
  • 40μl 1M formaldehyde

Continue incubation for 2hrs at 4°C

Then add

  • 10μl 1M dimethylamine-borane complex (ABC)

Incubate O/N at 4°C

Remove precipitated protein (up to 50%) by centrifugation

Concentrate and put protein on Gel Filtration column in a Tris containing buffer, eg:

  • 20mM Tris (pH7.5)
  • 200mM NaCl
  • 1mM DTT