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Dear Jahan,
is your diffraction useless, or do you call 4A resolution useless? 4A is
not too bad and can be a start for structure determination while you are
waiting for better crystals.
You give very little information about what you actually have d
Jahan
It sounds as though the protein crystallizes well, so microseeding (done
the right way) is very likely to solve the problem. Make a strong seed
stock with as much crystalline material as possible from several wells,
including different hits if possible. Just mix them all together, but keep
apart from optimising the crystallisation conditions, it might be
worth optimising the protein preparation or do some limited
proteolysis - or even express short N-terminal and/or C-terminal
deletions.
Mark
Quoting Patrick Shaw Stewart:
Jahan
It sounds as though the protein crystallizes
Dear CCP4 Users,
A CCP4 update has just been released, consisting of the following changes:
* PHASER: update to 2.5.2
* ctruncate: correction to twinning tests for H3
* DiffractionImage: further corrections to displaying of pilatus mini-cbf
(Linux and Mac)
* ccp4i: New ph
You might get better help if you post links (not attachments) to diffraction
images from several angles and an image of the crystal that gave the image, and
maybe several images of typical crystals.
Also, you could post dimensions of the crystal, and if your lab is set up for
it, a picture of t
Would someone suggest a cryoprotectant for index screen #59? It contains
0.02 M MgCl2
0.1 M HEPES pH 7.5
22% polyacrylic acid 5100, Na salt
Adding glycerol tends to dissolve the crystals.
Thanks,
Ed
Dear Ed,
try ca. 30 % PEG 400
Another possibility could be Perfluoropolyether Cryo Oil from Hampton Research.
This liquid doesn't mix with water. You put the crystal in few microliter and
remove the excess water droplets from your screen solution (otherwise you will
get ice rings).
Cheers,
Ul