Hi all, I have been trying to purify cytosolic fraction of membrane protein whose domain boundries are unknown. hence I have made a series of deletion constructs. The expression and purification is not a problem. I get good yields of the proteins. But on a gelfiltration column, they run in the void volume, which suggest that they are huge aggregates. I have tried different salts, DTT bME etc but no change.
I use sarkosyl in my lysis buffer. But could any one tell me if I can use any detergents as (bOD) in the purification protocol, and how much should I use. with regards Anita
