Dear Artem,
Thanks for your reply. You raise a number of points.
Immediately, I should comment that the price of the PX Scanner is very
considerably less than the $400k that you mention.
Whilst - with the proteins and crystallisation conditions that you may be
working
with, visual
Dear Marcus,
I always feel a bit uneasy about the advertisement-like posts that
Agilent (and others) place on this BB. Of course, there are
interactions between users and suppliers on many fronts, not least the
support you guys provide in the form of sponsorship to meetings and
OK,
I took the challenge. I got 7 out of 10. The three I missed were 2
questions about multi-well crystals which would be better (no problem)
and the capillary (no problem either, because you can mount it)...
I wouldn't be that snipe and braging (pun intended) if I would not
agree with
Dear All,
Here at NE-CAT, we make extensive use of XDS in a parallel environment. We are
looking to purchase some new hardware, so I am soliciting your opinions.
Our current cluster is made up of 16 nodes, each with 2 processors that have
four cores, running at 2.2 GHz (I believe). We run with
Hi Marcus,
I think you misinterpret my comments :) My first point is not that my
specific proteins make it easy to distinguish salt from protein visually -
it's rather that I am experienced enough and also know enough chemistry to
make an educated conclusion about the contents of my drops. I
Hi Frank,
the following are some recommendation for increasing the processing
speed of XDS. You can find them (and add to them !) at
http://strucbio.biologie.uni-konstanz.de/xdswiki/index.php/Performance .
Only item 7 is specific for a cluster.
In the order of effect:
1. XDS scales well
Hi Frank,
sorry, my first response was not very specific for your situation!
I studied your list and graphs and would just like to point out that in
2.1 Testing Filesystem Performance you may be severely overcommitting
the CPU resources, if this was for one machine only with 16 (=8+8)
cores.
Dear Artem,
Thanks for your very comprehensive reply. At the risk of reaping further
criticism,
I will now do little more than just reiterate my earlier comment: that from the
feedback
that we receive, the PX Scanner is much valued by the number and range of groups
that are now using these
Posted on behalf of Vertex Pharmaceuticals (please do not reply directly to
me!)
Research Scientist – Structural Biology
Vertex Pharmaceuticals Incorporated is a global biotechnology company
committed to the discovery and development of breakthrough small-molecule
drugs for serious diseases.
Hi all,
I am trying to decide between using a N-term his-tag or an N-term flag tag to
be expressed on a protein that I will eventually want to try and crystallize.
I usually don't cleave my tags unless absolutely necessary and I want to avoid
double tagging. I am leaning towards the flag
Dear CCP4 community,
I have a question a bit off CCP4 topic, but that could use so expert input.
While discussing about a bacterial secreted hemophore (heme scavenging
protein) for which the apo form has been solved, it seems that attempts to
obtain the Heme bound form are failing; in fact during
His-tag is close to neutral but flag-tag is acidic.
Using flag-tag for affinity purification usually gives cleaner protein but
it costs a fortune due to low binding capacity of commercially available
anti-flag resins.
--Chun
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On
On 04/19/11 14:18, Stephane Richard wrote:
Dear CCP4 community,
I have a question a bit off CCP4 topic, but that could use so expert input.
While discussing about a bacterial secreted hemophore (heme scavenging
protein) for which the apo form has been solved, it seems that attempts to
obtain the
Thanks everyone for your comments. The overall opinion appears to be that
large-scale purification of flag-tag protein is expensive. I 100% agree and
have thought about that plus the dilemma with the flag-tag elution difficulty
(low pH etc) which some of you also pointed out. So I will
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
My vote is for His-tag *unless* your protein is of the same general class as
Zn-fingers, B-boxes, etc. (weak/multisite/uncertain metal binders) - these
proteins often do not fare too well with His-tags because a) the tag
residues can participate in artefactual metal-binding sites and b) the use
of
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