Amylose pull-down of glycogen-bound proteins
From Bridges Lab Protocols
Revision as of 16:12, 7 May 2012 by Irith (Talk | contribs) (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
