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

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CCP4bb <-- 2008 <-- February 2008 <-- 19 February 2008
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Subject: FW: CCP4 web-newsletter 47 (Winter 2007-2008)
From: "Remacle, F (Francois)" f {- dot -} remacle {- at -} DL {- dot -} AC {- dot -} UK
Date: 2008-02-19 		Next message:
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Date: 2008-02-19


Subject: Re: crashing-out protein eluted from Nickel column
From: Sophia Tsai sophiats {- at -} USC {- dot -} EDU
Date: 2008-02-19
Hi,

Agreed on this. I used to have issues with aggregation due to the Nickel
being stripped off the column (happens with elution in imidazole). Adding
10mM EDTA to the elution immediately AFTER it has come out of the column
will chelate the Ni++ and prevents aggregation (at least in my case).
Afterwards, I use gel filtration to remove the Ni++, as well.

Hope that helps!
Sophia

On Mon, Feb 18, 2008 at 4:48 AM, Ngo Duc Tri wrote:

> Hi,
> I used another way to deal with this problem. You can try to elute your
> protein with the buffer containing 50mM EDTA (You need at least 10CV to
> elute completely). Then use gel filtration to remove the Ni.
>
> I applied this method with two proteins and it showed good results.
> Good luck!
>
> TriNgo
>
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