dear sanjiv
i dont think that the removal of detergent is more difficult than urea,since u
are purifying ur protein over Ni-NTA column,so after a few wash over there u
can remove the detergent completely.
best
atul kumar
-----Original Message-----
From: CCP4 bulletin board on behalf of David Cobessi
Sent: Tue 5/5/2009 6:18 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Refolding of Denatured Protein
Sanjiv Kumar wrote:
> I have not tried purifying this protein either with detergent or with
> /Guanidine Hydrochloride. I can try with both /Guanidine HCl and
> detergent. Since I got good purification with the 8M urea, I was
> thinking if this protein could be refolded and used further. Isn't
> removal of detergent more difficult then Urea?
> Sanjiv Kumar
> Lab. No. 411,
> Functional Genomics Unit,
> Institute of Genomics and Integrative Biology,
> New Delhi-110007
> India
Is it a transmembrane protein or not? or just a protein associated to
the membrane (with a GPI anchor for example)?
If it is a transmembrane protein, you should try to use detergents for
extraction and during the purification.
David
--
David Cobessi
Institut de Biologie Structurale
41, Rue Jules Horowitz
38027 Grenoble Cedex-1, France
Tel:33(0)438789613
33(0)608164340
Fax:33(0)438785122
