hi Andy
Here's my tip: If the protein is not denatured, but just precipitated, you can
use the precipitate as research tool, trying to see if the protein will
dissolve again with additives such as EDTA or divalent metal ions, or a bit of
salt, or cofactor ATP or NAD, detergent etc (anything non-denaturing)
This will learn you about what the protein likes, and helps you to make a
better prep that doesn't precipitate. We saw some miracles. If its rare protein
and if it dissolves again you can even try to go ahead as if nothing happened,
best
Joost
Dr. Joost Uitdehaag
Dept. Molecular Pharmacology
group leader protein crystallography
Schering-Plough Research Institute, Oss
tel: +31-412-666738 fax: +31-412-662519
-Original Message-
From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Juergen Bosch
Sent: Thursday, 02 October, 2008 4:11
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] resuspending precipitated protein
6M Guanidiniumhydrochloride works pretty good, the question is only
will you be able to refold it correctly ?
I believe a better approach is to avoid precipitation in the first
place and optimize your purification procedure in such a way that it
works. When do you observe precipitate ? Dialysis, then us e a fast
desalting column instead.
Jürgen
On 1 Oct 2008, at 06:12, ANDY DODDS wrote:
Hello,
following on from a previous topic about precipitating protein, but I
believe a distinct caveat of this warranting a separate thread, I
would like to know people's experiences in trying to get precipitated
protein back into solution? Is there a way or are there many ways?
Any experiences would be welcome,
yours,
Andy
-
Jürgen Bosch
University of Washington
Dept. of Biochemistry, K-426
1705 NE Pacific Street
Seattle, WA 98195
Box 357742
Phone: +1-206-616-4510
FAX: +1-206-685-7002
Web: http://faculty.washington.edu/jbosch
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