Re: [ccp4bb] finicky protein

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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Subject: REMINDER: Job opportunities- Helmholtz Department of Structural Biology, DESY Hamburg.
From: Christine Bentz Christine {- dot -} Bentz {- at -} HELMHOLTZ-HZI {- dot -} DE
Date: 2008-03-03

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Subject: Re: Fwd: Re: Glu+Arg solutions for solubility
From: Stephen Prince steve {- dot -} prince {- at -} MANCHESTER {- dot -} AC {- dot -} UK
Date: 2008-03-03

Subject: Re: finicky protein
From: Brenda Patterson ucbccka {- at -} UCL {- dot -} AC {- dot -} UK
Date: 2008-03-03

Did you filter your lysate through .45 then .22 filters?

cheers
b

Quoting James Stroud :

> 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 <-- 1999 <-- November 1999 <-- 30 November 1999

Next message:
Subject: Re: Fwd: Re: Glu+Arg solutions for solubility
From: Stephen Prince steve {- dot -} prince {- at -} MANCHESTER {- dot -} AC {- dot -} UK
Date: 2008-03-03