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Re: [ccp4bb] crashing-out protein eluted from Nickel column
 

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CCP4bb <-- 2008 <-- February 2008 <-- 15 February 2008
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Subject: crashing-out protein eluted from Nickel column
From: Juliana Barbosa Coitinho julianabbc {- at -} YAHOO {- dot -} COM {- dot -} BR
Date: 2008-02-15 		Next message:
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From: Bart Hazes bart {- dot -} hazes {- at -} UALBERTA {- dot -} CA
Date: 2008-02-15


Subject: Re: crashing-out protein eluted from Nickel column
From: Juergen Bosch jbosch {- at -} U {- dot -} WASHINGTON {- dot -} EDU
Date: 2008-02-15
Not sure if somebody mentioned it already, but have you played with
other buffers than the recommended sodium phosphate from the manual ?

Most lazy people use Tris because there's a 1 M stock solution - but you
can try also other buffers as long as you stay in a pH range >pH6 Of course you still need imidazole for the elution but the buffer might
bring some surprises, one of them can be a better initial purity after
the first capture step, another one might stabilize your protein.

Juergen


--

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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