Re: [ccp4bb] Protein DNA complexes concentration "bis"

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

Basic tutorials:

   - Chemistry
   - Protein
   - Peptide
   - Amino Acids

Xtal community:

- CCP4BB

CCP4bb navigation

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

Previous message:
Subject: Re: Positions / CCP4 wiki
From: Gerard DVD Kleywegt gerard {- at -} XRAY {- dot -} BMC {- dot -} UU {- dot -} SE
Date: 2008-10-07

Next message:
Subject: Computational Biochemistry Faculty Position Available
From: Jeff Watson watsonj {- at -} GMAIL {- dot -} COM
Date: 2008-10-07

Subject: Re: Protein DNA complexes concentration "bis"
From: Allyn Schoeffler aschoef {- at -} BERKELEY {- dot -} EDU
Date: 2008-10-07

We have had success concentrating protein by dialyzing the sample against
buffer plus 10-15% high molecular weight PEG (8-10k). It takes some trial
and error to get the right concentration; sometimes it helps to do a
stepwise procedure, gradually increasing the amount of PEG. Hope this
helps!

Best Regards,
Allyn

> Dear all
>
> For crystallisation assays and other experiments, I want to concentrate
> 2 protein-DNA complexes purified by gel filtration.
>
> - The affinity of the protein for the DNA is 10 nM.
> - I could concentrate the protein until 1.5 mg/ml (= 38 µM)
> - I have already tried lyophylisation (before and after complex
> creation) or amicon method without results (I can't manage to increase
> the concentration).
> - The complex could be a dimer or a tetramer on the DNA
>
> Do you have any good idea about this?????
>
> Thank you for your help and sorry for my first incomprehensible mail
>
> CHAIX Denis
>
> Centre de Biochimie Structurale
> INSERM U554/CNRS 5048
> 29 rue de Navacelles
> F-34090 Montpellier, France
> Tel: 33 4 67 41 77 21
> Fax: 33 4 67 41 79 13
> e-mail:Denis.chaix@cbs.cnrs.fr
> http://www.cbs.cnrs.fr
>

--
Allyn J. Schoeffler
Berger Lab
Dept. of Molecular and Cell Biology
UC Berkeley
phone: (510) 643-9491

CCP4bb navigation

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

Previous message:
Subject: Re: Positions / CCP4 wiki
From: Gerard DVD Kleywegt gerard {- at -} XRAY {- dot -} BMC {- dot -} UU {- dot -} SE
Date: 2008-10-07

Next message:
Subject: Computational Biochemistry Faculty Position Available
From: Jeff Watson watsonj {- at -} GMAIL {- dot -} COM
Date: 2008-10-07