Protocol Online logo
Top : Forum Archives: : Protein and Proteomics

TCA/Acetone precipitation - (Aug/11/2006 )

hi,

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?
Thx

-anitacindy-

you can try chilling the acetone and spinning it down or you can evaporate the acetone and resuspend the residue in sample buffer.

-mdfenko-

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.

-Missele-

Why do the proteins stay in the acetone if it's not shilled down? What does cooling down of the acetone actually does?

-smoochiepie79-