Forgot to hit the "Reply all" button (sorry :-( )
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Katja Schleider wrote:
Hi everybody,
sorry for my off-topic question. I got small initial crystals in 200mM
NaSulfat and 20% PEG 3350.
There is no buffer in this condition. How can I optimize these
crystals? Just vary the PEG concentration? Or should I add a buffer;
or vary the pH of the buffer the proteinsolution was in?
Thank you and best regards,
Katja
Hi Katja,
What we usually do here (IBS/LBM) is optimisation manually in 24 wells
plates. Usually pH vs precipitant concentration (for this you need a
buffer). Since you have crystals already, I would try to see if varying
the precipitant is enough (who knows, you might be lucky). If the
crystals we obtain are not of sufficient quality, we go on with the 3
additive screens from Hampton Research, also manually (droplets of 2
microl + 2 microl + 0.5 microl additive). This (additive screen) can be
done by robotics too.
Fred.
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