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
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
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
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
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
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
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
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,
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
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