I would like to thank all of you who promptly replied to my posting with so
many ideas and suggestions (18 answers so far). I will post a summary soon.
best wishes to all
Savvas
On Apr 19, 2011, at 12:48 PM, Savvas Savvides wrote:
Dear colleagues
We are working on a large bacterial
I would think thing here is that this protein actually associates to
those lipid nanodiscs...(around the disc) and Na cholate CMC is
around 10 mM.
so, yes you can solubilise proteins that bind lipids, the question is
does this protein bind lipids or not? or is it just scrambled or
Dear colleagues
We are working on a large bacterial protein (featuring a large number of
repeats) that appears to copurify with a lot of other proteins after
Ni-affinity chromatography and gel-filtration. We have tried adjusting the
ionic strength of these runs and have gone to as high as 5M
I recently followed a protocol from Stephen Sligar's lab for the purification
of his nanodisc protein, which has strong hydrophobic character as it
associates with phospholipids. His protocol includes washes with 1% Triton
X-100 and then with 50 mM cholate (both at pH 8 in the presence of 300
HI Savvas,
We recently had a protein that showed two overlapping peaks on the disposable
fast flow Q columns, so we decided to see if we could resolve them with a
higher resolution Q media. It ended up having 7 distinct peaks, only one of
which was free of contaminants. We have also noticed
