This might be obvious, but make sure you washed the crystals
thoroughly before dissolving them for SDS-PAGE or mass spec
-Jesse
On Thu, Jan 26, 2012 at 10:48 AM, Katherine Sippel
wrote:
> Might I suggest consulting the CCP4 user community wiki on the topic:
>
> http://strucbio.biologie.uni-konst
1. Try room temperature mounts (as suggested by others)
2. Expose the hell out of the crystal (5 min) on home source or go synchrotron
3. Run your protein through another column (ion exchange) even if it looks pure
4. Try an additive screen
5. Try limited proteolysis or methylation
6. If none of
Maybe, you can adjust the ion strength of your condition.
On Thu, Jan 26, 2012 at 8:32 AM, Kevin Jin wrote:
> For crystallization:
>
> Your xtal may come out a little bit fast. If the condition contain
> alcohol, such as IPA, you may have to modify it.
>
> If you let people know the condition
For crystallization:
Your xtal may come out a little bit fast. If the condition contain
alcohol, such as IPA, you may have to modify it.
If you let people know the condition, it may be more helpful.
Also, please check the purity of your protein.
Kevin
On Thu, Jan 26, 2012 at 7:33 AM, Theresa
Theresa
You should also try microseeding into *random screens *by making a seed
stock with the crystals that you have, to use with both microbatch and
vapor diffusion experiments. You will often pick up new and better
conditions and you're more likely to get well-formed crystals right out of
the
As other have/surely will suggest, by all means
try RT collection to establish if your crystals really diffract
OK, or if your cryo conditions are killing them. We have lots of
experience in our lab getting "beamstop" diffraction with certain
samples when subjecte
It depends a lot on which home source and which synchrotron, there are
enormous differences. Goettingen is uniquely well placed because we can
reach four synchrotrons in a few (3-7) hours by high speed train and in
theory at least five more with a longer train journey, trains are very
convenien
Ditto to Poul's advice.
I've had many many many cases where crystals diffract poorly (or not at all) on
home sources only to show excellent diffraction at a synchrotron. (Whether or
not a home source is properly calibrated is probably the biggest issue, but
that's for another discussion).
: eddie.snell Email: esn...@hwi.buffalo.edu
Telepathy: 42.2 GHz
Heisenberg was probably here!
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Poul
Nissen
Sent: Thursday, January 26, 2012 10:50 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] No diffraction
But
But first of all: try to add a synchrotron to the crystals
Poul
On 26/01/2012, at 16.48, Katherine Sippel wrote:
> Might I suggest consulting the CCP4 user community wiki on the topic:
>
> http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Improving_crystal_quality
>
> Good luck,
>
>
Might I suggest consulting the CCP4 user community wiki on the topic:
http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Improving_crystal_quality
Good luck,
Katherine
On Thu, Jan 26, 2012 at 9:33 AM, Theresa H. Hsu wrote:
> Dear crystallographers
>
> I have a protein of 90 kDa formi
Dear crystallographers
I have a protein of 90 kDa forming dimers. Crystals formed with microbatch and
vapor diffusion method in 24 hours but no diffraction at home source. Dissolved
crystals was confirmed to be the protein with mass spec.
Any suggestions to improve diffraction would be welcome.
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