Hi all.
I don't have any experience with Clontech and Fisher
resins, but about the GE one I faced the same problem as
Sebastiano indicated.
But I have to say that the problem was not general, but
protein (or family of protein) related: the low binding
depends on the oligomerization state of the chimeric
protein. When it behaves as dimer (GST dimerizes) it's
fine, when the oligomeric state increases (in my case
hexamer) I really can't manage to purify the protein on
GST sepharose. GST not exposed...I guess...
I'm waiting for the replies Mirek's question...I'm
interested too.
Ciao,
Barbara
On Wed, 5 Jun 2013 09:32:28 +0200
Sebastiano Pasqualato <[email protected]>
wrote:
Hi Mirek, hi all,
I'm also very interested in the topic, so please keep me
up with the replies, or make sure to post a summary,
please.
In addition to the price, the problem we're facing with
GSH-beads from GE (although we haven't tried others yet)
is that we can't manage to deplete our lysates.
We are always left with a large amount of unbound
GST-tagged protein in the flow through, that is
eventually captured by a second, third and sometime
fourth incubation with fresh beads.
Using larger beads volume won't help.
Has anybody faced and/or overcame this problem?
Thanks a lot in advance,
ciao,
Sebastiano
On Jun 4, 2013, at 8:54 PM, "Cygler, Miroslaw"
<[email protected]> wrote:
Hi,
I would like to ask the bb faithful for their experience
with the glutathione affinity resins. We have been using
so far the Glutathione Sepharose fast flow from GE but
the price is getting steeper. We found Glutathione
Superflow resin from Clontech to be significantly less
expensive and Glutathione agarose from Fisher somewhere
in-between. We have no experience with the latter two
resins and I wonder what is the experience of other
people with these resins? Do they have decent binding
capacity? Can they be efficiently regenerated or are they
a single use only?
Thanks for your help,
Mirek
--
Sebastiano Pasqualato, PhD
Crystallography Unit
Department of Experimental Oncology
European Institute of Oncology
IFOM-IEO Campus
via Adamello, 16
20139 - Milano
Italy
tel +39 02 9437 5167
fax +39 02 9437 5990
Barbara Giabbai
Elettra-Sincrotrone Trieste S.C.p.A.
SS 14 - km 163,5 AREA Science Park
34149 Basovizza, Trieste ITALY
Office: +39 040 375 8840
Lab: +39 040 375 8537
