a.
Add
1/10 volume of 100% TCA (w/v, made fresh) to solution to be precipitated and
vortex for 15 sec.
b.
Place on ice for a
minimum of 15 min.
c.
Centrifuge at 14,000
x g for 10 min.
d.
Remove and discard
the supernatant.
e.
Wash the pellet 2x
with 100 ul of acetone, by adding acetone, mixing, and then spinning for 5
min. Remove and discard the
acetone from the loose pellet.
Allow the final pellet to air dry thoroughly. The presence of residual acetone will make the resuspension
more difficult.
f.
Add desired volume
of 2x protein loading buffer and resuspend by vigorous vortexing.