membrane protein purification - solubilization and detergent removal (Feb/28/2005 )
Hi guys !
I am wroking on a membrane protein,from baculovirus expressed cells.The protein was in insoluble form and soluble only in the presence of strong anoinic detegents like SDS,Sarkosyl but not in others.It has enzymatic acticy also.
How to remove SDS from protein , and confirm my protein is active with out /less chage in structure ?
Apart form Kits which removed detergenys i9s there any way to remove anionic detergent?
-bonami_rv-
SDS , I beleive, can be removed by gel filtration or dialysis.
-sharath-
QUOTE (bonami_rv @ Feb 28 2005, 07:50 AM)
Hi guys !
I am wroking on a membrane protein,from baculovirus expressed cells.The protein was in insoluble form and soluble only in the presence of strong anoinic detegents like SDS,Sarkosyl but not in others.It has enzymatic acticy also.
How to remove SDS from protein , and confirm my protein is active with out /less chage in structure ?
Apart form Kits which removed detergenys i9s there any way to remove anionic detergent?
I am wroking on a membrane protein,from baculovirus expressed cells.The protein was in insoluble form and soluble only in the presence of strong anoinic detegents like SDS,Sarkosyl but not in others.It has enzymatic acticy also.
How to remove SDS from protein , and confirm my protein is active with out /less chage in structure ?
Apart form Kits which removed detergenys i9s there any way to remove anionic detergent?
You will never get rid of SDS totally. No matter what some biotech company tells you their product will do. There have been volumes written about it.
-mjray-