TCA/Acetone precipitation - (Aug/11/2006 )
After TCA precipitation of my sample, I spin down and saw a pellet. But when I washed my pellet with acetone, I couldn't see anymore my pellet. I removed the acetone (and kept it), put laemmli buffer in my eppendorf and test if my protein. I couldn't find it. I think my protein are still in the acetone. Does anyone have an idea how I can recover them?
you can try chilling the acetone and spinning it down or you can evaporate the acetone and resuspend the residue in sample buffer.
did you use cold acetone (-20°C) ?
if not, your protein can be in the acetone.
so do as mdfenko suggested : chill the acetone to -20°C, the proteins should precipitate.
Why do the proteins stay in the acetone if it's not shilled down? What does cooling down of the acetone actually does?