Resin is at 4deg, shake bottle well to homogenize slurry
Add 1ml of slurry to a 2ml tube
Spin at 2000 g 1-2 min
Wash 1ml of lysis buffer 3x
on last wash, remove buffer and add 1ml of purified protein
Rotate at 4°C for 1hr
Spin at 2000 g 1 min at room temperature
Remove supernatant and refreeze (safe to thaw 2-3 times)
Wash 1 ml of lysis buffer 3x
Resuspend in 1ml of wash buffer
Store beads at 4 deg, they are good for 2-3 months
Lysis Buffer
20mM HEPES pH7.5
10mM MgCl2
50mM KCl
1% Triton X-100
Add before use, final concentrations:
100ug/ml Cyclohexamide
20mM N-ethyl maleimide
1X Protease Inhibitor cocktail
1mM PMSF
1X Superase-in (Ambion)