[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

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

Previous message:
Subject: Re: Wanted: Structure with Lots of Model Bias
From: Pavel Afonine PAfonine {- at -} LBL {- dot -} GOV
Date: 2010-01-07

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From: Panne Daniel panne {- at -} EMBL {- dot -} FR
Date: 2010-01-08

Subject: Reg. Protein purification
From: Sivaraman Padavattan s {- dot -} padavattan {- at -} GMAIL {- dot -} COM
Date: 2010-01-08

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

Previous message:
Subject: Re: Wanted: Structure with Lots of Model Bias
From: Pavel Afonine PAfonine {- at -} LBL {- dot -} GOV
Date: 2010-01-07