To ask a potentially daft question - but why do you need to reduce the salt?
You say you are able to purify it, and that it behaves on gel filtration - but
only starts misbehaving when you dialyse, and the NaCl is below 100 mM.
We've crystallised many proteins, particularly DNA-binding
Dear Pascal,
We have the case of a protein that gelifies without precipitating when
bearking overexpressing E. coli cells. We had a look at the sample in
electron microscopy and the protein polymerises and forms irreversible
and very pretty fibers (explaining the gel). Maybe you want to have
Hi Pascal,
Is this a membrane protein? I worked on a GPCR that gelled upon
high concentration in a detergent/cholesterol solution. It was
reversible upon dilution and ran as a monomer by size exclusion
chromatography. Detergent assays showed that detergent was not being
concentrated in the
Dear All,
I am presently faced with a peculiar case in the lab. We are expressing a
protein in E. coli and we are able to express it as a fusion
protein without problems . Fusion cleavage goes well and the final product
looks homogenous by size-exclusion chromatography with the expected
molecular
reminds me of structure 1NEU, although here the gelation was reversible, see
ref and abstract below - the paper has a photo of a tube of soluble protein and
gelled protein
we have also had a couple of cold-sensitive proteins in our hands, that
precipitated at 4 degrees when concentrated, but
- Original Message -
From: Pascal Egea pas...@msg.ucsf.edu
To: CCP4BB@JISCMAIL.AC.UK
Sent: Monday, April 22, 2013 3:36:24 PM GMT -08:00 US/Canada Pacific
Subject: [ccp4bb] gelification of a pure protein
Dear All,
I am presently faced with a peculiar case in the lab. We
Anthony Duff Telephone: 02 9717 3493 Mob: 0431891076
-Original Message-
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Michael
Thompson
Sent: Tuesday, 23 April 2013 9:44 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] gelification of a pure protein
Does
Thanks to All for the diligent answers to my query,
The protein is not thermophilic and has only one cysteine. We are working
in presence of freshly added reducing agent and glycerol to promote
solubility (well kinda it seems).
This is not an RNA or DNA binding protein and it has no
Hi Pascal,
Your problem brings to mind this paper:
FG-Rich Repeats of Nuclear Pore Proteins Form a Three-Dimensional Meshwork
with Hydrogel-Like Properties Science 3 November 2006: 314 (5800), 815-817.[DOI:
10.1126/science.1132516]
Of course, in that case, the gellation was deliberate, but
