Amylose pull-down of glycogen-bound proteins
From Bridges Lab Protocols
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
