western and trizol - western and trizol (Dec/27/2007 )
Hi, anyone here did western blot using proteins from trizol treated cells?
From the instruction of trizol reagent (invitrogen), they said when you completed lysising the cells, the organic phase could be saved for protein precipitation. Wash the protein with guanidine hydrochloride and redissolve in 1%SDS, then the protein are ready for western blot.
I did once, but seemed not so good. Is it because this protocol not suitable for phosphorylated protein detection? I really need the RNA and the protein from the cells at the same time.
Ask for your kind help.
well i did once the protein precipitation from phenol phase.
I dissolved them in RIPA buffer, but the dissolution wasn't that good.
The total protein resuspention may not be possible so it's considered not a very good point to do this way.
Best is to prepare extra cells and split them in 2 : extract RNAs with one aliquot and proteins on the other one.
you may also consider the possibility to compare your protocol of protein extraction with the trizol-one and see if profiles are same.