[ccp4bb] crashing-out protein eluted from Nickel column

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CCP4bb <-- 2008 <-- February 2008 <-- 15 February 2008

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From: mesters mesters {- at -} BIOCHEM {- dot -} UNI-LUEBECK {- dot -} DE
Date: 2008-02-15

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Subject: Re: crashing-out protein eluted from Nickel column
From: Juergen Bosch jbosch {- at -} U {- dot -} WASHINGTON {- dot -} EDU
Date: 2008-02-15

Subject: crashing-out protein eluted from Nickel column
From: Juliana Barbosa Coitinho julianabbc {- at -} YAHOO {- dot -} COM {- dot -} BR
Date: 2008-02-15

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

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CCP4bb navigation

CCP4bb <-- 2008 <-- February 2008 <-- 15 February 2008

Previous message:
Subject: practical protein crystallization courses
From: mesters mesters {- at -} BIOCHEM {- dot -} UNI-LUEBECK {- dot -} DE
Date: 2008-02-15

Next message:
Subject: Re: crashing-out protein eluted from Nickel column
From: Juergen Bosch jbosch {- at -} U {- dot -} WASHINGTON {- dot -} EDU
Date: 2008-02-15