Hi,
In addition to the tips already suggested, you could also try to elute
your protein with histidine.. I've got a protein that crashes out with
imidazole as well, and I have succesfully used 200mM histidine for
elution (using either KPi or MES buffer in my case).
Cheers,
Ronnie
On Feb 15, 2008, at 5:47 PM, Juliana Barbosa Coitinho wrote:
Dear all,
I am with the same problem of Jacob.
My protein is precipitating, especially when I 'freeze it'. I am
using a His Trap HP column coupled with a desalting column. Already
tried to elute with gradient of imidazole (that allowed an elution
with less amount of imidazole), but even then, the protein still
crashes-out.
I am trying to use the protein immediately after purified it, but
this is not always possible and I am losing much protein with that.
I will try to use the tips that have already been said.
But if someone can help me more, I will thank!!!
Thanks a lot!
Juliana
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Ronnie Berntsson
----------
Ph.D. Student
Department of Biochemistry
Groningen Biomolecular Sciences and Biotechnology Institute
& Zernike Institute for Advanced Materials
University of Groningen
Nijenborgh 4, 9747 AG
Groningen, The Netherlands
telephone: +31 50 363 4195
telefax: +31 50 363 4165
e-mail: [EMAIL PROTECTED]
homepage: http://www.rug.nl/gbb/research/researchgroups/enzymology/index
