Dear ALL;
As an alternative strategy to avoid endotoxin, I plan to express the
protein in mammalian cells.
As suggested by others, the typical vector is pcDNA3.1(+). Does anyone
have comments on this vector or recommend some other powerful vectors?
I am new to mammalian exp
Dear ALL;
Thanks a lot for all the instructive suggestions. As my first trial, I
tried 0.1%Triton X-114 plus Ni-column binding buffer.
However, the oligomer of my protein got dissociated in to monomers as
indicated by gel-filtration.
Does anyone know how to resc
Min,
I forgot one more thing.
If you try to grow crystall of membrane protein and get tiny crystal,
you may try to add 5~10% IPA + 5mM NH4SO4 to addjuct the surface
charge of your protein.
The crystall may get larger. Folks like this trick.
Good luck.
Kevin
On Tue, Mar 13, 2012 at 10:38
Hi Min,
Please try this way if you use your protein for crystallization.
1. collect the needle and run SDS page or FPLC to verify the presence
of protein. Make sure it is not a buffer salt.
2. You don't need to do dialysis to remove b-ME, otherwise it will
take too long and you may lose some pr
The Nolen lab in the Institute of Molecular Biology at the University of Oregon
is seeking a highly motivated postdoctoral researcher to join our lab full time
on a flexible start date. Our interest is in understanding the molecular basis
for the regulation of the cytoskeleton. We are currently
I would try molrep - it will detect the NC translation and use it..
Eleanor
On Mar 13 2012, vincent Chaptal wrote:
Dear ccp4,
I have a case of PTS and wonder what's the best strategy to handle my data.
I processed my data in C2 with a=161 b=109 c=225 beta=104. The data in
97% complete to 3,
rwcontents xyzin my.pdb
Eleanor
It will not be able to guess the number og H atoms associated with the RNA
so the answer will be a bit out.
Eleanor
On Mar 12 2012, james09 pruza wrote:
Dear CCP4bbers,
Is there any tool to calculate the Matthews coefficient from a
crystallographic model o
That difference density looks too good to be true..
There is obviously a very strong non-crystallographic translation.
Is the spacegroup and/or cell correct?
Eleanor
On Mar 11 2012, xiaoyazi2008 wrote:
Hi All,
I have an interesting thing to share. 2.3A dataset with good quality, P21
Par
There are 3 options:
1) Use molrep. If you have two copies related with PST then it can give right
solution
2) Take the latest version of phaser and use it. It can deal with PST with two
related copies also
3) Reduce cell dimensions (it does not seem to be possible in your case. PST is
not exac
Dear Vincent,
The easiest thing for you to do right now in the CCP4 context is to run Molrep,
with the flag for PTS turned on. If that works and gives a clear solution,
that's great. If not, then I'd suggest downloading a recent version of Phenix
and trying the new version of Phaser that hand
Dear ccp4,
I have a case of PTS and wonder what's the best strategy to handle my data.
I processed my data in C2 with a=161 b=109 c=225 beta=104. The data in
97% complete to 3,8A.
xtriage detected a 40% peak in the patterson at fractional coordinates
x=-0,001 y=0,055 z=0,5.
I want to try to
Hello Deng,
Process them as usual with e.g. Mosflm, then sort them together (you
may need to rebatch one of the runs) and do one Scala run - this will
put all of the measurement on a common scale and write out data
suitable for refinement. For pointgroups with ambiguous origin choices
(e.g. P4) yo
Hi CCP4bb,
I have two dataset from one crystal with different distance.now i am wondering
how to merge the two dataset into one file,and use it to refinement.
thanks a lot!
deng
Using an envelope or a mask to start phasing even with high NCS is still worse
than using a low res model from cryoEM that is characterized by an (almost)
proper density distribution.
One has to measure the intensities of the very low res reflections (0 0 1 and
the likes) precisely.
The presen
Mosflm works fine with our Pilatus 2M at DLS - however you do need to
be using the beta-test version.
http://www.mrc-lmb.cam.ac.uk/harry/mosflm/betas/
best wishes,
Graeme
On 12 March 2012 19:57, Dean Derbyshire wrote:
> Hi again, it's the 2M detector I'm having problems with. Got different d
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