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Created page with '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 i...'
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
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
