[Protein-crystallography] Re: help! membrane protein solubilization

Marc Böhm maboehm01 at arcor.de
Thu Jun 9 16:42:50 EST 2005


aumium wrote:
> Hi, friends.
> 
> I have a membrane protein which consists of 12 TMs. I expressed it in
> E. coli and isolated the membrane fraction. Then I tried to extract it
> from the membrane with different detergent, but just failed even with
> 1%SDS.
> 
> Could anybody give me some clue? Thanks.
> 

Hi!

Did you tried also additional parameters like pH, salts, Temperature?

By the way, are you sure that your protein is properly integrated into 
the membrane. I had a target which remained in inclusion bodies inside 
the cell... even the strongest detergent would fail if the protein isn't 
part of the membrane fraction.



Regards

Marc




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