[ccp4bb] very high concentration of protein

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

Previous message:
Subject: 2010 Gordon Research Conference on Diffraction Methods
From: Andrew Leslie andrew {- at -} MRC-LMB {- dot -} CAM {- dot -} AC {- dot -} UK
Date: 2009-07-02

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Subject: Re: multi-domain protein with identical tertiary structure
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2009-07-02

Subject: very high concentration of protein
From: peter hudson peter {- dot -} hudson {- dot -} peter {- at -} GMAIL {- dot -} COM
Date: 2009-07-02

Hello all

I am working with a small protein-protein complex. This complex express
quite well . I purify in a buffer of pH=9.0 with 150mM NaCl and 1% of
glycerol and able to concentrate upto 20 mg per ml. I have a two clones of
this protein complex. One is N-terminal His tagged and another C-terminal
His-tagged. While concentration of the N-terminal His tagged protein in
cnetricon it forms yellow color slimy deposition on the surface of membrane.
while C-terminal His tagged protein does form very highly viscous layere at
the surface of membrane but it is completly colourless.I aliquate the
concentrated protein by pippetting into different aliquate rather than
collection of whole protein by centrifugation. if i check the concentration
of the last aliqoute(which isbottommost viscous protein part) both prtoein
complex, it shows very high concentration compared to the first fraction. I
have not done DLS.
Is my both C-terminal His tagged tagged as well as N-terminal His tagged
protein are forming soluble aggregates.

I would appreciate the help.

Thanks in advance

Peter

CCP4bb navigation

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

Previous message:
Subject: 2010 Gordon Research Conference on Diffraction Methods
From: Andrew Leslie andrew {- at -} MRC-LMB {- dot -} CAM {- dot -} AC {- dot -} UK
Date: 2009-07-02

Next message:
Subject: Re: multi-domain protein with identical tertiary structure
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2009-07-02