Path: utzoo!utgpu!watmath!iuvax!bionet!primeb.dundee.ac.uk!BI0712
From: BI0712@primeb.dundee.ac.uk
Newsgroups: bionet.molbio.methds-reagnts
Subject: Solubilization of inclusion bodies
Message-ID: <8904112209.AA05688@net.bio.net>
Date: 12 Apr 89 04:53:59 GMT
Sender: daemon@NET.BIO.NET
Lines: 19


Can anyone help me solubilize my protein from inclusion bodies? Having
overexpressed my bacterial peripheral membrane protein I am having
a difficult time getting the inclusion bodies into solution. I have tried
the standard denaturants 8M urea and 5M guanidium with a variety of
addatives eg DTT, mercaptoethanol, triton, tween, NaCl without much luck.
The protein will go up in 8m urea 20mM MES [C [C [C [C [C [C [C [C [C if heated
 to 65C for
10 mins, but I don't imagine I will be able to refold a protein subjected
to that sort of treatment.
Anybody had these sort of problems, or got any ideas [D [D [D [D [D  [C [C [C [C
 [C [C [C got any ideas?                                                   [D [D
Any help gratefully received.

Steve Hyde
Molecular Genetics
Dept of Biochemistry
University of Dundee
Dundee. DD1 4HN. UK.