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

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CCP4bb <-- 2008 <-- February 2008 <-- 14 February 2008
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Subject: Re: rescuing crashing-out protein eluted from Nickel column
From: Chun Luo chun {- at -} ACCELAGEN {- dot -} COM
Date: 2008-02-14 		Next message:
Subject: Re: rescuing crashing-out protein eluted from Nickel column
From: José_Trincão trincao {- at -} DQ {- dot -} FCT {- dot -} UNL {- dot -} PT
Date: 2008-02-15


Subject: Re: rescuing crashing-out protein eluted from Nickel column
From: Jacob Wong jacob {- dot -} j {- dot -} wong {- at -} GMAIL {- dot -} COM
Date: 2008-02-14
Thank you all for the quick replies - I'm trying many of the tips in small
batches and would send out a feedback as soon as appropriate, gratefully, -
J.J.

On 2/14/08, Jacob Wong wrote:
>
> Dear all,
>
> I just ran into this problem and would like to see if I could get some
> helpful tips before my protein completely crashes out.
>
> I have a protein as 6His fusion and it remained bound to the Ni resin with
> 40 mM Imidazole wash (added to 1XPBS) but then was eluted off with 200 mM
> (added to 1XPBS). The protein seemed to be highly concentrated in the
> elution and began to get cloudy right away, with more and more precipitation
> produced over a matter of minutes. I felt so helpless, didn't know what to
> do, and then decided to add 5% of glycerol into one of the fractions but
> that made it even more cloudy (ohh no...).
>
>
> While the protein is dying in the tube, do you have some quick remedy for
> me? Thanks very much, -J.J.
>
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