Amylose pull-down of glycogen-bound proteins

Protocol

  • All steps on ice/cold room
  • Add 50 ul beads (amylose resin #E80021L Biolabs) to 0.5 mg of protein lysate in lysis buffer with protease inhibitors
  • Add 1 ml of lysis buffer
  • Rotate in cold room 30’ to ON
  • Spin down at max speed for 1’
  • Aspirate supernatant down to 1 mm over beads with crushed tip.
  • Repeat 3-5 washes with lysis buffer without protease inhibitors
  • After last wash and aspiration spin again and aspirate all supernatant
  • Ad 50ul SDS loading buffer, boil for 5 minutes
  • Load 10ul/gel