I've found that crystallization in sitting drops under oil dramatically reduces
the no. of nucleation events and increases the overall crystal size. Now, if
the increased crystal size helps improve diffraction is something to be tested.
Among the many other suggestions...
Raji
-Includ
Not a contradiction at all :) If the protein supply is adequate - this is
a perfectly good approach to explore. I simply forgot to mention it.
I've also not mentioned engineering the protein's surface (works really
well for us because we have secret ways to do this, learned from ancient
and myster
In addition to trying lower concentrations of PEG and protein, you might
consider whether aggregation in your protein stock or vibration in your
crystallization environment are contributing to the problem, and how
they might be minimized. It might be obvious, but we have found that it
is important
You said you tried macroseeding, but what about microseeding? It works
quite well in many cases. Just don't forget that you should find a zone
were nucleation does not happen before you seed. Take a look at this paper:
Bergfors, T. (2003) Seeds to Crystals, J. Struct. Biol., 142, 66-76.
There
Hi Jobi
Sounds like you need to explore your protein vs PEG concentration, and my
guess (directly contradicting Artem's) is that chances are you need to pump
[protein] way up and drop [PEG] way down.
Like in [protein]=>30-40mg/ml, [PEG]<=2-4%. Often people don't go into that
range when setti
What often works for this scenario:
Change temperature. Try 21, 15, and 4C.
Add glycerol or ethylene glycol - set a screen with your condition as basis,
and % EG/glycerol across - from 1% to 15% in suitable increments
Find additives that produce no crystals, set up tray(s) with those, then
se
