On 20/04/2020 20:23, Kyle Gregory wrote:
I am assessing ligand binding and each of the monomers display density at the site but it is not as clear as
I would like. [] I was wondering if it is feasible, or if there are any
tools, that can be used to improve density based of the fact there are t
I take it that you have 2 molecules in the asymmetric unit. If so, you could try some sort of NCS averaging of the map or just NCS-refinement might help. Not much more here than has been expertly suggested already. Can you give us an idea of the resolution? How many RMS have you contoured the map a
It sounds like you are looking for MAPROT.
http://www.ccp4.ac.uk/html/maprot.html
On 2020-04-20 15:23, Kyle Gregory wrote:
Hello all,
I have a homodimer structure in P1 21 1 spacegroup, the dimer is
likely a crystallographic artefact, where it looks like the monomer is
rotated by ~ 180 degr
Hmmm - your space group imposes a too fold screw acid parallel to the Y
axis? If your dimer 2-fold is also parallel to the Y axis you should see a
strong non crystallographic translation vector? Is that correct? Such
things can complicate density. Eleanor
On Mon, 20 Apr 2020 at 20:24, Kyle Grego
Hello all,
I have a homodimer structure in P1 21 1 spacegroup, the dimer is likely a
crystallographic artefact, where it looks like the monomer is rotated by ~ 180
degrees around the Y axis.
I am assessing ligand binding and each of the monomers display density at the
site but it is not as cle
If your protein is His-tagged: spin down cells @~1500xg, adust pH with tris
buffer and NaOH to ~pH 7.1 (spin again higher speed if necessary to remove more
particulates) and pass over chemical resistant Ni-resin.
Linda Olson, PhD
Assistant Professor/
x-Ray Facility Manager
Dept. Biochemistry
Med
Gentle spin followed by TFF - a cheap peristaltic pump plus $200 filter
cartridge will do the trick.
Artem
On Mon, Apr 20, 2020, 11:56 AM Gloria Borgstahl
wrote:
> Hi Friends, We are secreting Spike Ecto domain into the media from insect
> cells for purification. As we scale up I am wondering
I used to do a low g centrifugation (100g for about 10min), collect the
supernatant and filter it onto Polycap units (Whatman) 0,8µm/0,2µm (the size
will depend on your volume) using a large peristaltic pump (Masterflex).
> Le 20 avr. 2020 à 17:56, Gloria Borgstahl a écrit :
>
> Hi Friends, W
Hi Friends, We are secreting Spike Ecto domain into the media from insect
cells for purification. As we scale up I am wondering what is recommended
for collecting the media from large volumes of culture. Centrifugation?
Filtration of some kind? I imagine we need to be gentle to not lyse the
ins
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