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Something else to try would be the Protic Ionic Liquid kit from Hampton.
I recently had crystals of a protein that would only grow as laminated
stacks of plates. Optimizing the conditions and using an additive screen
didn't improve crystal morphology. I tried the PIL kit from Hampton and
was able to get single, thick plates in several conditions. At the ACA
meeting in Hawaii a few years back, there was a poster about ageing your
PEG solutions by microwaving them and letting them cool on the bench.
This was the only way that the poster's author could get reproducible
crystals of her target protein. 

 

Good Luck!

Bryan

 

From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of
Regina Kettering
Sent: Wednesday, August 24, 2011 2:47 PM
To: [log in to unmask]
Subject: Re: [ccp4bb] spherulites and PEG3350

 

Something to consider is the quality of the PEG 3350.  We have found
that different qualities of PEG 3350 can give different results,
depending on the type and amount of contaminants.  What used to be the
Fluka PEG 3350 is now the pharm grade of PEG 3350 (aka Miralax).  We use
high quality PEG 3350 for normal screening, but switch to the highest
quality grade we can get for optimizing.

 

Regina

 

________________________________

From: Jan van Agthoven <[log in to unmask]>
To: [log in to unmask]
Sent: Wednesday, August 24, 2011 2:05 PM
Subject: [ccp4bb] spherulites and PEG3350

Dear all,

 

I recently obtained some spherulites while trying to crystallize my
protein. The spherulites are manually reproducible, but changing pH,
protein concentration, and salt concentration does not result in crystal
formation. Microseeding with crushed spherulites isn't a solution either
as it only yields new spherulites. Next stepp is the use of an
optimization kit but I have a limited amount of material, and I start
doubting that these are protein spherulites, as the spherulites are not
particularly soft. The condition contains 15% PEG 3350 and 200 mM NaCl.
Does anyone know if PEG 3350 forms easily spherulites around that
concentration?

 

 

Thanks, 

 


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