Re: [ccp4bb] finicky protein

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Subject: Re: finicky protein
From: Tim Gruene tg {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2008-03-03

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From: Christine Bentz Christine {- dot -} Bentz {- at -} HELMHOLTZ-HZI {- dot -} DE
Date: 2008-03-03

Subject: Re: finicky protein
From: James Stroud jstroud {- at -} MBI {- dot -} UCLA {- dot -} EDU
Date: 2008-03-03

Try refolding before purification.

On Mar 2, 2008, at 11:47 PM, Tim Gruene wrote:
>> Hi all
>>
>> sorry, for offtopic query...
>>
>> I am trying to purify my protein by Ni-NTA affinity chromatography.
>> After
>> sonication as i centrifuge bacterial lysate, soon after 10 min
>> whole lysates
>> get precipitated during loading on the column and some time it remain
>> soluble too. if i get purified through the column without
>> precipitation, it
>> gets precipitated during dialysis.
>> I have tried lot, by chnaging buffers, increasing salt or
>> deacreasing salt
>> or no salt at are helpless.
>> I do purifiaction in cold room.
>>
>> can any one suggest some solution?
>>
>> Thanks in advance.
>>
>> NSH
>>

--
James Stroud
UCLA-DOE Institute for Genomics and Proteomics
Box 951570
Los Angeles, CA 90095

http://www.jamesstroud.com

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CCP4bb <-- 2008 <-- March 2008 <-- 03 March 2008

Previous message:
Subject: Re: finicky protein
From: Tim Gruene tg {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2008-03-03