[ccp4bb] finicky protein

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From: Khan Amir Rafiq amirrafk {- at -} TCD {- dot -} IE
Date: 2008-03-02

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Subject: Re: finicky protein
From: Tim Gruene tg {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2008-03-03

Subject: finicky protein
From: Naren sharma narensharma11 {- at -} GMAIL {- dot -} COM
Date: 2008-03-02

Hi all

sorry, for offtopic query...

I am trying to purify my protein by Ni-NTA affinity chromatography. After
sonication as i centrifuge bacterial lysate, soon after 10 min whole lysates
get precipitated during loading on the column and some time it remain
soluble too. if i get purified through the column without precipitation, it
gets precipitated during dialysis.
I have tried lot, by chnaging buffers, increasing salt or deacreasing salt
or no salt at are helpless.
I do purifiaction in cold room.

can any one suggest some solution?

Thanks in advance.

NSH

CCP4bb navigation

CCP4bb <-- 2008 <-- March 2008 <-- 02 March 2008

Previous message:
Subject: PDF, PhD and technician positions in Ireland
From: Khan Amir Rafiq amirrafk {- at -} TCD {- dot -} IE
Date: 2008-03-02

Next message:
Subject: Re: finicky protein
From: Tim Gruene tg {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2008-03-03