Dear all,
Thanks for the many of responses, the data from Crysalis is scaled, but
unmerged so needed to be fed through scala/truncate before running Phaser.
Phaser is now running with no problems and I am looking at some nice maps.
Bestr wishes and thanks again for your help,
Steve
Dr
Dear all,
Can I draw your attention to the below post doctoral scientist opportunity at
Diamond.
I24 is the microfocus MX beamline at Diamond. We are looking for a PDRA to
build on recent exciting results in the fields of radiation damage, fast data
collection and in-situ spectroscopy. The
Hi all,
Has anyone performed protein peptide interaction experiments using
flurophore on a real time PCR machine?
Please advice me on how the experiment is performed. Can I use ANS?
with regards
--
rashmi
Yes
add peptide in different concentrations to your protein and run the assay*
Yes, or Sypro Orange
*Rule of thumb 20 kDa protein use 1 mg/ml 50 kDa protein use 0.5 mg/ml
also have a peptide which should not interact with your protein as control
Jürgen
On Jun 28, 2012, at 7:34 AM, rashmi
I can second this. Whether or not you get a signal with DSF depends on your
protein, not on the type of ligand. We also use Sypro orange.
Herman
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of
Bosch, Juergen
Sent:
Dear crystallographers
Trying to crystallize a membrane protein complex of 100 kDa with a soluble
protein of 20 kDa which is interact with the membrane protein. So far, no
co-crystals in 200 conditions. Some conditions gave crystals but mass spec of
crystals show only either one protein
I would advise against using anti-His FaBs for that particular purpose.
Even if you get them to bind to your complex, the inherent flexibility of
the terminal regions were His-tags are placed might ruin the final result,
and they're not cheap to begin with.
You can further characterize your
Try seeding the complex with the crystals that you have. If you have
crystals of both proteins, crush them and mix them together. Suspend
the seeds in whichever reservoir solution has the least salt in it*
(or suspend in 50% PEG**) . Use random Microseed Matrix Screening
(rMMS) i.e.
Postdoctoral Research Associate--Biochemistry and Structural Biology of
Microtubule Nucleation
A post-doctoral position is available in Dr. Ken Sawin’s laboratory, in the
Wellcome Trust Centre for Cell Biology, University of Edinburgh, UK to study
the molecular mechanisms of microtubule
In trying to reproduce a very nice public structure a cloning mis-hap put a
-GS- prior to the C-term 6His tag. The resultant crystals had a 500Ang C
dimension and 16 molecules in the au. Even a single amino acid could make all
the difference
David Hargreaves
Associate Principal Scientist
There are plenty of reports (some even successful) where small concentrations
of proteases have been used to enable crystallisation (with soluble proteins).
Having put the effort into getting your material in the first place it's a
small extra step to try. The professional way is to run a time
Dear All,
Thanks for all the replies on and off-board. I received around twenty replies
and the majority have spoken in favor of the QIAgen BSA-free anti-5His mAb from
QIAGEN. Not to be bias, a couple of people recommended the one from Abcam as
well.
Thanks again, Dan
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