An alternative could be to disolve your cholesterol in methanol or maybe in
50% methanol, then add it directly to the drop. Does your protein tolerate
10-20% methanol? Then you can add the methanol-cholesterol solution you
your hanging drops directly and mix fast. The methanol is volatile and will
Jerry,
Steroids have a certain solubility also in ethylene glycol and PEG.
You might be able to work out a crystallization strategy around this.
The ethanol/PEG combination was used in:
http://www.nature.com/nature/journal/v437/n7055/full/nature03923.html
Enrico.
Dear ALL:
Sorry for
if you are crystallizing membrane protein
here is a useful protocol from JCIPMT website
follow the link
http://jcimpt.scripps.edu/protocols/JCIMPT_PreparationofCHSStock.pdf
in my experience even after extensive sonication i filter the buffers
otherwise we found that cholestrol comes out of
Dear ALL:
Sorry for this kind of off-topic question.
I am going to co-crystallize one protein with cholesterol. I read some
papers saying that
their protein can be pre-incubated with 1mM cholesterol in the presence of 5%
(v/v) ethanol.
TO do so, I first dissolved cholesterol in
