Try refolding before purification.
On Mar 2, 2008, at 11:47 PM, Tim Gruene wrote:
Hi all
sorry, for offtopic query...
I am trying to purify my protein by Ni-NTA affinity chromatography.
After
sonication as i centrifuge bacterial lysate, soon after 10 min
whole lysates
get precipitated
Dear colleagues,
we are writing to remind you that 6 March 2008 is the closing date for
applications for the following job opportunities.
The German Helmholtz Association, together with the University of
Hamburg, is creating a new department of Structural Biology at the
German Electron
Did you filter your lysate through .45 then .22 filters?
cheers
b
Quoting James Stroud [EMAIL PROTECTED]:
Try refolding before purification.
On Mar 2, 2008, at 11:47 PM, Tim Gruene wrote:
Hi all
sorry, for offtopic query...
I am trying to purify my protein by Ni-NTA affinity
Hi folks
We are pleased to announce the release of Mosflm version 7.0.3 and
iMosflm 0.6.1.
These are bug-fix releases for versions 7.0.2 and 0.6.0, which affect
the following detectors and circumstances only. If your detector is not
listed, it isn't affected, so you shouldn't need to update
I will second this recommendation to use alternate cell disruption
methods instead of sonication.
Particularly with multi-protein complexes, where the preservation of the
complex from disruption though
crystallization is important, sonication can be quite deleterious.
Even with the French
Dear CCP4 experts,
I'm working on a kind of cystein peptidase. It shows a little degradation
after 2 days storing at 4 degree even I tried many kinds of protease
inhibitors, or glycerol and DTT.
So I decided to work fast (1 day) then set xtal. After one week I pick up
some condition and check
Hi all,
I had an interesting experience, and wonder if others have seen
similar things.
I was collecting data from a crystal that contains an iodinated
macromolecule. After 2 days on a copper rotating anode, with the
crystal at 100 K, we experienced a detector problem, so I put the
Ngo Duc Tri wrote:
Dear CCP4 experts,
I'm working on a kind of cystein peptidase. It shows a little
degradation after 2 days storing at 4 degree even I tried many kinds
of protease inhibitors, or glycerol and DTT.
So I decided to work fast (1 day) then set xtal. After one week I pick
up some
If your protein is autolyzing, you may need to crystallize it in the
presence of an inhibitor. In the case of trypsin, I have crystallized
in the presence of benzamidine, then removed the inhibitor by dialysis
after crystallization.
kmj
Ngo Duc Tri wrote:
Dear CCP4 experts,
I'm working on
Yes indeed ! H2 is one product of H2O radiolysis by recombination of two H
radicals.
Whether or not the cleavage of iodine (quite efficient under X-rays) is a
factor increasing H2 production, I don't know.
Philippe Dumas
IBMC-CNRS, UPR9002
15, rue Rene Descartes 67084 Strasbourg cedex
tel: +33
RESEARCH POSITION IN THE ROSSJOHN LABORATORY
http://www.med.monash.edu.au/biochem/staff/rossjohn-lab.html
The protein crystallography unit at Monash University, headed by Prof
Jamie Rossjohn, ARC Federation Fellow, seeks two talented scientists to
join his group.
One position to work in
I used glutaraldehyde vapor by placing crystal drop above a bridge
with 15% glutaradehyde for various amount of time. It worked for me
for many crystals, but not for the one that I am trying now. The
crystal didn't diffract any more. I intended to make the crystal
tougher for heavy metal soaking.
Dear structural(-ly interested) biologist !
HIC-Up, the Hetero-compound Information Centre - Uppsala, has been updated and
now contains information on 7,870 hetero-entities. This is the first HIC-Up
version after the release of the remediated PDB. A few changes had to be made
to the
Dear Crystallographers:
just to share something I found out recently, that is really helpful to
know:
5 min Copper Stain protocol
Normal SDS-PAGE gels can be stained in 5 min with 300mM CuCl2. Simply remove
the gel, rinse ~10s in water, place 5 min in CuCl2, and the gel is
negatively
One down side is that copper sulfate is toxic to fish and plants. Please
be good to the environment and do not pour copper sulfate solutions down
the drain. We normally precipitate with NaOH, filter, and put the
precipitated copper in an appropriate waste.
-Original Message-
From:
There's an even faster reagent for staining - takes abut 1 minute overall
and the results look just like dear old Coomassie. This reagent works
wonderfully for us. Yes, the recipe is proprietary - but if you add up the
cost of reagents for normal Coomassie and compare - this one isn't much more
Touche'
Jacob
***
Jacob Pearson Keller
Northwestern University
Medical Scientist Training Program
Dallos Laboratory
F. Searle 1-240
2240 Campus Drive
Evanston IL 60208
lab: 847.491.2438
cel: 773.608.9185
email: [EMAIL PROTECTED]
There's an even faster reagent for staining - takes abut 1 minute overall
and the results look just like dear old Coomassie. This reagent works
wonderfully for us. Yes, the recipe is proprietary - but if you add up the
cost of reagents for normal Coomassie and compare - this one isn't much more
Hello,
I did the replacement soaking and it was successful. Now I want to
superimpose the two maps to show (I hate to show off but I have to) the
map diffrence of the ligand region to demonstrate that the replacement
soaking was a success. Is there a way to superimpose the two maps?
The
Yanming Zhang wrote:
Hello,
I did the replacement soaking and it was successful. Now I want to
superimpose the two maps to show (I hate to show off but I have to) the
map diffrence of the ligand region to demonstrate that the replacement
soaking was a success. Is there a way to superimpose
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