If you spin the crystal through a full 360 degrees then any given relp
(unique reciprocal lattice point) will pass through the Ewald sphere no
more than twice. This is because the Ewald sphere is a surface and the
relp is taking a circular path. It is either inside the Ewald sphere
or
I am trying to crystallize a protein that I am phosphorylating in-vitro. There
is no way that I can purify the phosphorylated protein from unphosphorylated
one. I tried Ion exchange and gel filtration for separating them and they are
not working.
Mass spec result shows the phosphorylation at
Thank you Prof Lewis.
Its serine residue which is getting phosphorylated. the pI of my protein is
around 8.2. So native gel ( around pH 8.3) is also not working for me. I tried
ion exchange at pH 7.0, 7.5 and 8.5 but my protein is either coming in flow
through or very poorly binding to Q and SP
On 18/01/15 18:29, Kenneth A. Satyshur wrote:
Coot is our best program there ever was for fitting electron density. It is
very simple to
use and easy to teach. But sometimes improvements are made that seem to just
slow
things down. Having used 0.7.1 for a long time, i noticed that rotation and
Try running a regular SDS page gel w/o boiling your sample. Some P proteins run
differently on gel due to the conformational change that is induced. I agree
that you probably have a mixture in which the P form might be the minor
species. I'd probably try to get the IEX to work; given that the
Hi, there:
If you just want to know the percentage of phosphorylation, I highly recommend
phospho-tag gel.
You can buy the phospho-tag, and make gels your self. It's very effective.
Shawn
Toronto
On 2015-01-19, at 1:01 PM, Bert Van-Den-Berg wrote:
Try running a regular SDS page gel w/o
an effective modification to your protein preparations would be to add
phosphatase inhibitor tablets during all
stages of purification. Pierce sell a combination protease and PI tablet
that is effective and economical to be added during protein preps. Also you
invitro buffers should have
Dear All,
I checked the Mg2+ B factor, and it was around 22. So, were the values for F
atoms. For AlF3, it was more than 50 for Al and more than 40 for F atoms. I
will take care about the occupancy. I hope that will fix the small negative
density.
When I inserted MgF in COOT using the code MGF
Don't forget, multiplicity has its own negative connotations.
http://www.imdb.com/title/tt0117108/
Shane Caldwell
McGill University
On Sun, Jan 18, 2015 at 12:26 PM, Edward A. Berry ber...@upstate.edu
wrote:
Also RAID (REDUNDANT array of inexpensive disks). To me redundancy implies
