Dear ccp4
I
wanted to ask if anyone has expressed DDB1 (Damaged DNA Binding protein) or
any other interacting partner using
bacterial cells instead of insect or mammalian or yeast cells, I also wanted to
ask if
anyone has any experience with this binding partner interacting with a fusion
Hello
Does anyone has a vector with multiple cloning site followed by protease
cleavage and C-terminal MBP for expression in Bl21 cells.
Thank you
Geula
--
Geula Davidov
Lab Manager
The Zarivach laboratory, Building 39, Room 0013
The National Institute for Biotechnology in the Negev
Dear all,
Please find here an advertisement for a postdoctoral position at the
European Synchrotron Radiation Facility in Grenoble, for which the
knowledge of protein crystallography would be an asset:
http://esrf.profilsearch.com/recrute/fo_annonce_voir.php?id=232
The work will consist in
Hi Bernhard,
If your cheap imidazole works fine aside from the absorption problem, there's
always my favorite stone-age detection method: pencil a numbered grid onto a
piece of filter paper, spot the fractions on it, and stain with coomassie.
It'll tell you which fractions to load on a gel,
Hi Bernhard,
this one works for us:
Sigma BioUltra imidazole, 99.5% by GC, 56749
Cheers,
Jan
--
Jan Abendroth
Emerald BioStructures
Seattle / Bainbridge Island WA, USA
home: Jan.Abendroth_at_gmail.com
work: JAbendroth_at_embios.com
http://www.emeraldbiostructures.com
On Aug 21, 2013, at 7:33 AM,
Would you happen to know what the A280 of a 1M (or .1 M) solution is?
I wonder if this absorbance is really due to impurities or is the tail of a weak absorbance band of imidazole itself. I
notice A280 is not one of the properties sigma lists for its imidazole.
Jan wrote:
Hi Bernhard,
this
According to Qiagen
Since imidazole absorbs UV radiation at 280 nm, an elution profile
measured at 280 nm while purifying a 6xHis tagged protein by FPLC will
show an increase in absorbance above the background signal allowing
quantitation of your protein. The absorbance of imidazole can vary
Sigma Imidazole 56749.
http://www.sigmaaldrich.com/catalog/product/sigma/56749?lang=enregion=US
Zhen
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Bernhard
Rupp
Sent: Wednesday, August 21, 2013 10:33 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Low 280 absorbance
Bernard, Ed,
Sounds like an impurity, all right. But if so you wold think
the chemical companies would be selling special low-A280 grade
imidazole for IMAC, and for some price you could get material
that doesn't absorb there at all. Unless its a spontaneous
decomposition product. Maybe even the
How about low pH elution or EDTA as alternative ?
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
Phone:
Hi everybody,
I would just like to contribute that absorption at 280 results from
methylated imidazole, a contamination hard to get rid of. Imidazole
itself does not absorb at 280.
Bye,
Matthias
-
Dr. Matthias Zebisch
Division of Structural Biology,
Hi bb,
I have a question about TLS group assignment. I have a loop that is poorly
ordered. The backbone can still mostly be built, though that's probably
just the predominant conformation of many. TLSMD usually assigns a group
border on one or both sides of this loop, though the assignment isn't
Hi Shane,
Option one and two may both work. The extra group is not a problem unless it
doesn't actually improve the fit with the data much. If you really have very
little data, you should consider first using one overall residual B-factor
and try to get the most out of the TLS. You can try
Yes, I think that's the one we use now. I believe it is the current branding of
the Fluka imidazole that we always ordered. Definitely lower UV absorbance. We
also prefer Roche high purity DTT for the same reason.
John
Sent from my iPad
On Aug 21, 2013, at 11:04 AM, Jan
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