[ccp4bb] AW: [ccp4bb] Off topic: His-tag 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: Re: Off topic: His-tag purification
From: Gregory T Costakes gcostake {- at -} PURDUE {- dot -} EDU
Date: 2012-01-16

Next message:
Subject: La Caixa"/IRB Barcelona PhD fellowships Programme selection procedure
From: Esther Fernandez Rodriguez esther {- dot -} fernandez {- dot -} rodriguez {- at -} IRBBARCELONA {- dot -} ORG
Date: 2012-01-16

Subject: AW: Off topic: His-tag purification
From: "Metlitzky, Moritz, Dr {- dot -} " moritz {- dot -} metlitzky {- at -} HELMHOLTZ-MUENCHEN {- dot -} DE
Date: 2012-01-16

Hi Theresa,

did you try to change the position of the His-Tag?
Perhaps it is not exposed in your current construct and would work better at the N- or C-terminus.
Greetings
Moritz Metlitzky, Ph.D.
Institute of Toxicology
Tel.: 089-3187-2669
Fax: 089-3187-3449
www.helmholtz-muenchen.de/en/toxi/krappmann
________________________________
From: "Theresa H. Hsu"
To: CCP4BB@JISCMAIL.AC.UK
Sent: Sunday, January 15, 2012 1:23:33 PM
Subject: [ccp4bb] Off topic: His-tag purification

Hi all

I have a His-tagged soluble protein (8 His residues added to 90 kDa protein) that do not bind to IMAC column based on flowthrough showing up with Western blott. Do you have suggestions to improve the binding?

Binding condition is 50 mM Tris-HCl 8.0, 300 mM NaCl, 10 mM imidazole pH to 8.0.

Thank you.

Theresa

________________________________
Helmholtz Zentrum M?nchen
Deutsches Forschungszentrum f?r Gesundheit und Umwelt (GmbH)
Ingolst?dter Landstr. 1
85764 Neuherberg
www.helmholtz-muenchen.de
Aufsichtsratsvorsitzende: MinDir?in B?rbel Brumme-Bothe
Gesch?ftsf?hrer: Prof. Dr. G?nther Wess und Dr. Nikolaus Blum
Registergericht: Amtsgericht M?nchen HRB 6466
USt-IdNr: DE 129521671

CCP4bb navigation

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

Previous message:
Subject: Re: Off topic: His-tag purification
From: Gregory T Costakes gcostake {- at -} PURDUE {- dot -} EDU
Date: 2012-01-16