I have been using an HRP conjugated antibody from Thermo that can detect
maybe 3-8ng of HisTagged protein from insect cell lysates. Performance is
ok, but with the sensitivity I am not so happy with.
How is the sensitivity with the Qiagen antibody? How many picograms you
need for a clear band?
Our experience essentially agrees with QIAGEN's claim that you can detect
proteins in the low nanogram range, i.e. 1-2 ng... so not a major improvement
over your current antibody, I'm afraid. They also sell an HRP conjugate version:
Dear ALL,
The threonine is found in the active site of my protein structure
from the crystallization in the absence of any threonine containing
chemicals. I presume that's why there was no signals detected with the
ITC experiment in which I titrate my protein with Threonine. In the in
Dear All,
I would like to draw your attention to a post doctoral vacancy in my group,
located at the Division of Structural Biology (StruBi), University of Oxford.
Your role will be to study the structure of an enveloped virus using electron
cryo-microscopy (cryo-EM). Necessary training will
Hi Wenhua
Have you tried using 6-8M urea or guanidinium hydrochloride to denature
your protein+threonine, then refold by dialysis into a
non-threonine-containing buffer? If you are performing ITC, then i guess
you already have an efficient activity assay, thus maybe use that as an
indicator of
I don't know the exact reaction your protein is performing, but an
alternative would be to try to turnover your threonine substrate by
adding the other substrates. The reaction product should more easily
leave the active site. You might have to do an additional gelfiltration
to get rid of excess
I'll just second, third, and fourth the assessment of the Qiagen
anti-pentaHis antibody. I pretty much gave up on anti-His antibodies until
we tried this out in the lab and now everyone loves it. Reasonable
sensitivity and specificity (we use IR dye-conjugated secondaries and a
Li-Cor Odyssey
Dear ccp4 bulletin board
I apologise for off topic question. I wonder if anybody knows of a good method
to detect oligomerisation?
I suspect an equilibrium intermediate is forming oligomers based on tryptophan
fluorescence showing an exposed tryptophan becoming buried in a hydrophobic
region.
Native PAGE (i.e. BN-PAGE), light scattering (i.e. MALLS)
Not quick and easy but could work: AUC (i.e. a sedimentation equilibrium
experiment)
-Brad
On Tue, Jun 26, 2012 at 9:01 AM, Careina Edgooms
careinaedgo...@yahoo.comwrote:
Dear ccp4 bulletin board
I apologise for off topic question. I
Dear Wenhua,
in extension to the post from Herman, you could try to remove the
threonine with a separate enzyme, for instance threonine oxidase or
amino acid deaminase. Their reaction products probably also bind less
tightly. The question is, how fast the release of threonine from your
Dear All:
Are there any web sites that produce stunning artistic images from pdb
files?
david
--
David M. Mueller
Biochemistry and Molecular Biology
The Chicago Medical School
Rosalind Franklin University of Medicine and Science
Green Bay Road
North Chicago, IL 60064
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Follow the link and click on her website:
http://web.me.com/bosch_lab/jbosch/News/Entries/2012/4/7_Cover_figure_in_Journal_of_Structural_Biology_%28April_2012_issue%29.html
In case the link doesn't work then just go to
http://web.me.com/bosch_lab
and click on the News
Never mind my previous email, I misread - I thought Journal Cover figures.
What's wrong with PyMol or PovRay DoItYourself ?
Jürgen
On Jun 26, 2012, at 9:44 AM, David Mueller wrote:
Dear All:
Are there any web sites that produce stunning artistic images from pdb files?
david
--
David M.
This is to inform you that a well-respected member of the crystallographic
community has published a totally hilariously absurdly funny Sci-Fi novel.
I added a link on my web site, www.ruppweb.org , and no, I said 'well
respected', so I did not write it, but maybe you can guess from the
Dear CCP4bb
I have collected a data-set using the supernova x-ray generator from Agilent
and taken the mtz file generated by the data processing software in crysalis
pro forward for structure solution. The data collection was straight forward
and the software seemingly processed the data
Hi Ed,
Another way to look for point mutations are the sequence clusters available at
100%, 95%, 90%, etc. sequence identity.
Here is an example: find mutations for HIV-1 protease:
Start with an example protein structure, i.e., 1OHR. On the structure summary
page of 1OHR click on the Seq.
I apologise for off topic question. I wonder if anybody knows a good
buffer/condition for promega protev?
I need to remove his-tag from my sensitive mamalian protein which does not
like the NaCl concentration less than 300mM.
Best,
Jahan
Hi, all,
I thought I could easily find a reference to comment upon the relative
intensity of rings in an image due to diffraction by polycrystal ice, but
no luck googling for that. A reference that would contain a picture (with
visual relative intensities) would be even better. Of course
Don't know where the exact problem is. However, it is definitely possible to
use a Crysalis-Scala-Truncate-Phaser pipeline without runtime errors. I have
done a few times.
I am sure you will be able to get help from Agilent people. If not, feel free
to get back to me.
Best
Roberto
On 26 Jun
Maybe figure 4 in
Viatcheslav Berejnov et al. Vitrification of aqueous solutions
J. Appl. Cryst. (2006). 39, 244–251 ?
JORGE IULEK wrote:
Hi, all,
I thought I could easily find a reference to comment upon the relative
intensity of
rings in an image due to diffraction by polycrystal
I think an appropriate reference is Bragg (1921)
http://dx.doi.org/10.1088/1478-7814/34/1/322 who compared various
possible crystal structures to the relative strength of the
reflections from ice powder measured by Dennison (1921)
http://dx.doi.org/10.1103/PhysRev.17.20.
However, as Bragg noted
Google yields amongst others:
His-tag impact on structure
Acta Cryst. (2007). D63, 295301
...quäl dich.
-Original Message-
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of weliu
Sent: Tuesday, June 26, 2012 6:07 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] The
Protein Biophysics Laboratory
Several positions at postdoctoral, research associate and research assistant
levels are available now in Protein Biophysics Laboratory, National University
of Singapore.
The successful postdoctoral candidate should demonstrate a strong motivation in
scientific
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