Re: [ccp4bb] Reg. Protein purification

- 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: announcement EMBO Practical Course : Scientific Programming and Data Visualisation for Structural Biology
From: Panne Daniel panne {- at -} EMBL {- dot -} FR
Date: 2010-01-08

Next message:
Subject: Re: Cavity filling
From: Simon Kolstoe s {- dot -} kolstoe {- at -} UCL {- dot -} AC {- dot -} UK
Date: 2010-01-08

Subject: Re: Reg. Protein purification
From: Jurgen Bosch jubosch {- at -} JHSPH {- dot -} EDU
Date: 2010-01-08

How about using an anion exchanger after your NTA step ?

JÃ¼rgen

......................
JÃ¼rgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry & Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
Phone: +1-410-614-4742
Lab: +1-410-614-4894
Fax: +1-410-955-3655
http://web.mac.com/bosch_lab/

On Jan 8, 2010, at 7:41, Sivaraman Padavattan
wrote:

> Dear all,
> I am trying to express the human protein using bacterial expression
> strain (Rosetta) and purified using Ni-NTA affinity purification.
> The Molecular weight of out protein is 47 kDa. In SDS-PAGE, we have
> seen that 27 kDa contaminant protein co-eluted with our protein even
> at high concentration of Imidazole. In Superose 12 column, these two
> proteins eluted as a single peak and its corresponding molecular
> weight suggestive of partial interaction. By mass mapping we have
> found 27 kDa band is an adenylate kinase. Is there any specific way
> to separate adenylate kinase from our protein?
> Thanks in advance,
>
> Sivaraman Padavattan
>
>
>

CCP4bb navigation

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

Next message:
Subject: Re: Cavity filling
From: Simon Kolstoe s {- dot -} kolstoe {- at -} UCL {- dot -} AC {- dot -} UK
Date: 2010-01-08