Re: [ccp4bb] Tag, you're in! Flag, V5, etc

Protein crystallography Programs for crystallography X-ray detectors
Basic tutorials:
Protein Peptide

CCP4bb navigation

CCP4bb <-- 2008 <-- February 2008 <-- 07 February 2008

Previous message:
Subject: Re: Solved: PDB file column-cut-paste issues
From: Raji Edayathumangalam redayath {- at -} MAIL {- dot -} ROCKEFELLER {- dot -} EDU
Date: 2008-02-07

Next message:
Subject: Conference Announcement: Quo Vadis Structural Biology?
From: Ben Eisenbraun bene {- at -} CRYSTAL {- dot -} HARVARD {- dot -} EDU
Date: 2008-02-07

Subject: Re: Tag, you're in! Flag, V5, etc
From: David M Shechner shechner {- at -} MIT {- dot -} EDU
Date: 2008-02-07

> At least that's been our experience.
> Andy
> (No affiliation with NEB)

Hi, everyone:

Just to give both sides to this story, though, we've had phenomenally bad luck
with the NEB Intein system (tagged on either the C- or N- terminus). The
protein expressed and purified beautifully, and post-cleavage was
pretty much a
single band on a silver-stained gel. However, it was intractably misfolded,
while purification with other, more 'traditional' tags (His6 or - dare I say
it - *untagged* purification!) yielded fully soluble, folded protein. A few
other labs around MIT have had similar experiences. So, for any technique
there will be circumstantial data affirming or refuting its utility; take any
success or failure story with the required ug of NaCl. =)

Good luck purifying,
Dave

CCP4bb navigation

CCP4bb <-- 2008 <-- February 2008 <-- 07 February 2008

Previous message:
Subject: Re: Solved: PDB file column-cut-paste issues
From: Raji Edayathumangalam redayath {- at -} MAIL {- dot -} ROCKEFELLER {- dot -} EDU
Date: 2008-02-07

Next message:
Subject: Conference Announcement: Quo Vadis Structural Biology?
From: Ben Eisenbraun bene {- at -} CRYSTAL {- dot -} HARVARD {- dot -} EDU
Date: 2008-02-07