Re: [ccp4bb] Reg. Protein purification

- Protein crystallography

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

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Subject: Re: Blob help
From: Tim Gruene tg {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2010-01-08

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Subject: Re: Reg. Protein purification
From: Jacob Keller j-keller2 {- at -} MD {- dot -} NORTHWESTERN {- dot -} EDU
Date: 2010-01-08

Subject: Re: Reg. Protein purification
From: Quyen Hoang qqhoang {- at -} GMAIL {- dot -} COM
Date: 2010-01-08

I have seen this when expressing sub-domains of a larger protein.
Adding some (about 0.5% w/v) BOG (beta octylglucoside) worked well for
cases similar to what you've described.

Cheers,
Quyen

_______________________________
Quyen Hoang, Ph.D
Department of Biochemistry and Molecular Biology,
Stark Neurosciences Research Institute
Indiana University School of Medicine
635 Barnhill Drive, Room MS0013D
Indianapolis, Indiana 46202-5122

Phone: 317-274-4371
Fax: 317-274-4686
email: qqhoang@iupui.edu

On Jan 8, 2010, at 7:41 AM, 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
>
>
>

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

Previous message:
Subject: Re: Blob help
From: Tim Gruene tg {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2010-01-08

Next message:
Subject: Re: Reg. Protein purification
From: Jacob Keller j-keller2 {- at -} MD {- dot -} NORTHWESTERN {- dot -} EDU
Date: 2010-01-08