Dear Raji,
In our lab we always
(1) use TCEP
(2) high molar ratio of protease with gentle rocking at 4C O/N
(3) keep in mind the size (KDa) of the target protein regarding to the
detergent micelle size. Although has been observed that in many cases the use
of SUMO tags increases expression
Dear colleagues,
with this message I would like to announce our workshop entitled:
*"Strategic pipeline planning: from sample preparation to 3D structure
determination with bio SAXS and other biophysical techniques"*
to be held at the National Hellenic Research Foundation (NHRF) in Athens
from Apr
Hi Raji-
To echo what John Lee said, we found that we had to maintain a ratio
between 1:2 and 1:10 Ulp1:substrate for several different target proteins,
none of which were membrane proteins however. So, I would definitely try
maybe a 1:5 ratio and see what happens. This wasn't too cost prohibitive
Hi Folks,
Thanks very much for your helpful suggestions. Several folks have suggested
switching to TCEP. I am also going to increase the molar ratio of Ulp1 in
the cleavage reaction like John Lee has suggested since I did not think of
the effect of DDM on Ulp1. (Everything seemed to work in the ca
Dear CCP4 friends,
Sorry first for the out of topic request.
We are going to buy more crystal screening kit, however, we are only familiar
with some popular kits which we already have in the lab (listing below). Do you
have your favourite kits which can share with us? Our lab more focus on enz
Hi Raji,
I've used SUMO/Ulp1 for some of my membrane proteins. My experience is you
have to add a lot of protease. Sometimes, I add 1-1 molar ratio to get the
job done.
Just to add, I've experienced similar results as you in 0.1%DDM, partly
because Ulp1 precipitates in the detergent (worse in OG),
Dear all,
The biannual Gordon Research Conference in Structural Biology, accompanied by
the first Gordon Research Seminar, will take place in the last week of July at
Bates College, New England, a few hours drive from the IUCr meeting that
follows in the first week of August.
The theme for the
Dear Friends of the Twilight,
the annual update of the twilight data base is now available:
Download:
http://www.ruppweb.org/twilight/default.htm
Readme:
http://www.ruppweb.org/twilight/Readme.htm
Papers:
http://journals.iucr.org/d/issues/2013/02/00/issconts.html
http://journals
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Dear Lisa,
Good think is that your protein is crystallizing and diffracting X-rays. I am
not sure
if your protein is
Dear Lisa, because we know nothing about our object: protein is membrane
or globular? Is there known structures of homologs?
What buffer do you use?
17.02.2014 13:57, LISA пишет:
Dear All,
My proein is polymerzated and elutate in the void volum when runnning
gel filtration Superdex 200. I can
Hi folks
We are pleased to announce a new release of Mosflm & iMosflm; there
are quite a few changes (a non-exhaustive list is at the end of this
message). The default web-pages for both programs will now lead you
directly to the new versions;
http://www.mrc-lmb.cam.ac.uk/harry/im
Dear Lisa,
the first thing to check is whether the polymerization is due to disulfide bond
formation. If you run a gel of your protein with one sample boiled in the
presence of b-mercapto ethanol and one sample boiled in the absence of bME,
this should tell you whether disulfide links are the cu
Dear All,
My proein is polymerzated and elutate in the void volum when runnning gel
filtration Superdex 200. I can get a small crystal after a lot of
optimization. But the resolution is still very low (about 8A ). I try to
find the sites involved in polymerization. Is there some software to
predi
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