[ccp4bb] finicky protein

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: finicky protein
From: David M Shechner shechner {- at -} MIT {- dot -} EDU
Date: 2008-03-03

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Subject: Re: finicky protein
From: price {- at -} UCHICAGO {- dot -} EDU price {- at -} UCHICAGO {- dot -} EDU
Date: 2008-03-03

Subject: finicky protein
From: Mads Gabrielsen mg118m {- at -} UDCF {- dot -} GLA {- dot -} AC {- dot -} UK
Date: 2008-03-03

I am not a big fan of sonication. Try changing your way of disrupting the
cells.

I have compared sonication vs mechanical stress on several unrelated proteins,
and for me a good old french press wins every time. If you want to get all
modern and fancy, a cell disruptor gives similar results.

Cheers,

Mads Gabrielsen

[Hide Quoted Text]

On Mar 2, 2008, at 11:47 PM, Tim Gruene wrote:

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

--
Dr. Mads Gabrielsen

GBRC, B217
Division of Biochemistry and Molecular Biology
IBLS
University of Glasgow Phone Office: 01413308119
G12 8QQ Phone Lab: 01413306449
UK E-mail: m.gabrielsen@bio.gla.ac.uk

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CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: finicky protein
From: David M Shechner shechner {- at -} MIT {- dot -} EDU
Date: 2008-03-03

Next message:
Subject: Re: finicky protein
From: price {- at -} UCHICAGO {- dot -} EDU price {- at -} UCHICAGO {- dot -} EDU
Date: 2008-03-03