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Re: [ccp4bb] Protein-DNA complex prepartion for crystallization
- Protein crystallography

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999
Previous message:
Subject: CONECT records
From: Christina Bourne bournecr {- at -} YAHOO {- dot -} COM
Date: 2008-10-02		Next message:
Subject: Re: CONECT records
From: "Schubert, Carsten [PRDUS]" CSCHUBER {- at -} ITS {- dot -} JNJ {- dot -} COM
Date: 2008-10-02


Subject: Re: Protein-DNA complex prepartion for crystallization
From: Djordje Francuski francuz {- at -} CHEMIE {- dot -} FU-BERLIN {- dot -} DE
Date: 2008-10-02

Just wandering, is your DNA dissolved in a buffered solution or just water?
I would suggest to buffer your DNA solution prior mixing with the protein.
Otherwise I guess that your pH in the protein-DNA mix is lower then the
expected pH7.0 and maybe your protein does not like it.

Cheers,
Djordje


> Dear All
> Sorry for non-crystallography query. I am working on
> DNA binding protein, while mixing DNA with protein for
> preparing Protein-DNA complex for crystallization,
> protein is precipitating. pI of the protein is 9.3
> and in 15 mM HEPES 7.0, 150mM NaCl and 5% glycerol.
> Concentration of the protein used for mixing with DNA
> is 8 mg/mL. DNA to protein molar ratio is 1.2. Please
> advise me how to prevent precipitation. Is changing of
> buffer pH and adding divalent cation like MgCl2 can
> help in preventing precipitation?
> Thanking You
> Rajakumara
>
>
> E. Rajakumara
> Postdoctoral Fellow
> Strcutural Biology Program
> Memorial Sloan-Kettering Cancer Center
> New York-10021
> NY
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CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999
Previous message:
Subject: CONECT records
From: Christina Bourne bournecr {- at -} YAHOO {- dot -} COM
Date: 2008-10-02		Next message:
Subject: Re: CONECT records
From: "Schubert, Carsten [PRDUS]" CSCHUBER {- at -} ITS {- dot -} JNJ {- dot -} COM
Date: 2008-10-02
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