Often you can chemically crosslink the GST-fusion to the GSH-agarose
resin, thus immobilizing your fusion. The chemical crosslinking could
destroy the desired binding properties however if binding activity is
maintained it is a robust way to keep the GST-fusion out of your
eluted protein(s) of interest.
Mike
===================================================================
.-----. Michael Curtis
/ \-_-/ \ Scientist II
/_-_\ /_-_\ Bristol-Myers Squibb Company
----- ----- Biotechnology Development
\-_-/ \-_-/ E-Mail: Michael_S_Curtis@ccmail.bms.com
\ /_-_\ / Phone: 315-432-2209
`-----' Fax: 315-432-4785
===================================================================
______________________________ Reply Separator _________________________________
Subject: GST fusion protein
Author: Christoph Turck <turck@itsa.ucsf.edu> at *INTERNET*
Date: 5/28/99 3:21 PM
Like many other groups that are interested in protein-protein
interactions we are using GST fusion proteins bound to glutathione
resins to pull out proteins from cellular lysates that specifically bind
to our protein of interest. After absorption and extensive washing we are
confronted with the problem of how to elute the specifically bound
protein(s) from the GST fusion protein without eluting the latter from the
glutathione resin. Does anybody know how much salt, urea, guanidine-HCl,
etc. one can get away with for eluting the specifically-bound protein
without eluting the GST fusion protein itself? Any recommendations are
welcome.
Happy Holiday,
Chris (turck@itsa.ucsf.edu)
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Date: Fri, 28 May 1999 15:21:41 -0700 (PDT)
From: Christoph Turck <turck@itsa.ucsf.edu>
Subject: GST fusion protein
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