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

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CCP4bb <-- 2008 <-- February 2008 <-- 18 February 2008

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From: ºîº£·å houhf {- at -} IHEP {- dot -} AC {- dot -} CN
Date: 2008-02-18

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Date: 2008-02-18

Subject: Re: crashing-out protein eluted from Nickel column
From: Ronnie Berntsson r {- dot -} berntsson {- at -} RUG {- dot -} NL
Date: 2008-02-18

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
>
> Abra sua conta no Yahoo! Mail, o único sem limite de espaço para
> armazenamento!

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: r.berntsson@rug.nl
homepage: http://www.rug.nl/gbb/research/researchgroups/enzymology/index

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CCP4bb <-- 2008 <-- February 2008 <-- 18 February 2008

Previous message:
Subject: refinement problem of labelled RNA complex
From: ºîº£·å houhf {- at -} IHEP {- dot -} AC {- dot -} CN
Date: 2008-02-18

Next message:
Subject: Re: refinement problem of labelled RNA complex
From: Manish Chandra Pathak pmxtal {- at -} YAHOO {- dot -} CO {- dot -} IN
Date: 2008-02-18