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

Protein crystallography Programs for crystallography X-ray detectors
Basic tutorials:
Protein Peptide

CCP4bb navigation

CCP4bb <-- 2008 <-- February 2008 <-- 14 February 2008

Previous message:
Subject: BCA/CCP4 Summer School on Macromolecular Crystallography XV
From: Elspeth Garman elspeth {- at -} BIOP {- dot -} OX {- dot -} AC {- dot -} UK
Date: 2008-02-14

Next message:
Subject: Re: rescuing crashing-out protein eluted from Nickel column
From: Chun Luo chun {- at -} ACCELAGEN {- dot -} COM
Date: 2008-02-14

Subject: rescuing crashing-out protein eluted from Nickel column
From: Jacob Wong jacob {- dot -} j {- dot -} wong {- at -} GMAIL {- dot -} COM
Date: 2008-02-14

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.

CCP4bb navigation

CCP4bb <-- 2008 <-- February 2008 <-- 14 February 2008

Previous message:
Subject: BCA/CCP4 Summer School on Macromolecular Crystallography XV
From: Elspeth Garman elspeth {- at -} BIOP {- dot -} OX {- dot -} AC {- dot -} UK
Date: 2008-02-14

Next message:
Subject: Re: rescuing crashing-out protein eluted from Nickel column
From: Chun Luo chun {- at -} ACCELAGEN {- dot -} COM
Date: 2008-02-14