Re: [ccp4bb] Solubilization buffer

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

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Subject: Re: methews coefficient and validation of 60mer
From: Pankaj Chauhan pankajimtech {- at -} GMAIL {- dot -} COM
Date: 2009-12-02

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From: tommi kajander tommi {- dot -} kajander {- at -} HELSINKI {- dot -} FI
Date: 2009-12-03

Subject: Re: Solubilization buffer
From: Joerg Standfuss jst {- at -} MRC-LMB {- dot -} CAM {- dot -} AC {- dot -} UK
Date: 2009-12-03

Dear Meg,

There is a database of refolding conditions from which you could get some
inspiration.

http://refold.med.monash.edu.au/search.php

The Urea conditions I have seen there mostly use 37C. for
solubilization/refolding and nearly all contain a reducing agent. Have a
look. There are also a great number of other additives people use to
improve things.

Joerg

> Hi all,
>
> We use 8M urea solubilization buffer for our protein in inclusion bodies
> and
> recommended temperature is 10-15º C. but in 8M conc the urea does not
> dissolve and is in crystalline form only, will it have any effect on
> solubilzation efficiency. Our solubilization time is 1 Hr and after that
> we
> centrifuge and use the supernatant for refolding via dialysis. however the
> pellet after centrifugation of solubilzation show presence of our protein
> on
> sds page analysis. what should we do so that the process of solubilization
> is complete and our protein is not lost in pellet.
>
> thanks in anticipation.
>
> meg
>

CCP4bb navigation

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

Previous message:
Subject: Re: methews coefficient and validation of 60mer
From: Pankaj Chauhan pankajimtech {- at -} GMAIL {- dot -} COM
Date: 2009-12-02

Next message:
Subject: UV detection of crystals/survey
From: tommi kajander tommi {- dot -} kajander {- at -} HELSINKI {- dot -} FI
Date: 2009-12-03