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