Please join us live or register to view on-demand:
Machine Learning Approach to Cellular Cryo-Tomography
November 01, 2023
03:00 PM (London) 11:00 AM (New York) 08:00 AM (San Francisco)
Speakers:
Ellen Zhong, Princeton
Abhay Kotecha, Thermo Fisher Scientific
Hi Rafael, Dom et al,
Indeed nickel eluted with imidazole stays on all your proteins with a His tag,
and addition of DTT will make almost all proteins go brown and crap out at this
point. But, as Dom says, addition of EDTA before the DTT will solve the
problem. Most of the time…
If your
Dear CCP4 community,
I have open positions in my group at the Center for Structural Biology, Center
for Cancer Research, National Cancer Institute in Frederick, Maryland, USA.
Please can you share the following details with anyone you think may be
interested.
I’m currently looking for
Hi Rafael,
For completeness - there are alternatives to imidazole for eluting proteins
from Ni-NTA resins:
EDTA - (strips Ni2+, and therefore everything bound to the Ni2+) - 50mM should
be sufficient, in your favourite buffer/salt system. Obviously not appropriate
for metalloproteins. You'll
Hi Rafael,
Once I inherited a protocol with a problem similar to yours and they told me
that the precipitation was caused by nickel leaking out during the elution with
250 mM imidazole. I am not sure whether this was true, however, their fix was
to place something like 20 ul of 200 mM EDTA at
Dear all,
A NIH-funded postdoctoral research associate position is available at Dr.
Wei Zhang's laboratory at the University of Minnesota (Minneapolis,
Minnesota, USA). The successful candidate will study enveloped
virus assembly and entry using cryo-EM, single particle reconstruction,
cryo-ET,
Hi Rafael,
the simple answer is that the EDTA and DTT( or any other reducing
agent) is not required for the TEV activity.
You can have your TEV protease in 20 mM Tris pH 7.5 (RT) and 150 mM
NaCl, plus either 10% Glycerol or 50% glycerol depending on storage
conditions preference - 80 or -20C
Hi everyone,
I have been looking on this bb and other websites as well but I could not find
a veredict. We are suspecting that when I elute my sample from my Ni-NTA
column, the imidazole concentration (250 mM) is making it to precipitate. Once
my sample has a cleavable TEV site, I was planning
Hi All,
The next Industry Talk will be "Hydrogen bond donor-acceptor asymmetries in
drug design" given by Peter Kenny (retired) on 29 November 2023 at 2 pm UK
time. Details and free registration link can be found at
https://www.ccpbiosim.ac.uk/hydrogenbond2023.
Abstract: The presentation will
Dear Jorge,
as Harry mentioned, autoPROC [1] will automatically exclude image
ranges that are significantly worse than the rest (i.e. add mostly
noise). This could be due to radiation damage, badly centred crystals
or anything else. The so-called "fitness parameter" it uses takes
multiplicity and
Dear Daniel,
is this for a Bruker Diffractomete? If you have a choice, I would be
surprise, because Bruker refused to offer a
new D8 with an EIGER2R500 detector - they'd rather not sell their
system, and we got a STOE Stadivari instead.
We've had a PhotonIII for about 1/2 year on our D8. It is
Hi,
Well, it seems there are already many good indications to study and to
work on.
Thanks, Oliver, Kay, Harry and Graeme!
Now, brain and hands on!
Jorge
Forwarded Message
...
Dear all,
I have found many fundamental studies on image processing and
Along similar lines, if you happen to be working in the DIALS ecosystem there
is the dials.damage_analysis program, which takes the scaled data (scaled.expt,
scaled.refl) and computes a number of statistics including Rd as discussed
below, and the cumulative-pairwise R factor Rcp, which
Hi
I’ve never actually used it in anger (one should never be angry when processing
data…), but doesn’t AutoProc, developed by the good folks at Global Phasing do
a lot of these analyses? Clemens, Claus etc may have something pertinent to say.
Harry
> On 30 Oct 2023, at 13:23, Jorge Iulek
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