Re: [ccp4bb] how to make cholesterol solution

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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Subject: Re: how to make cholesterol solution
From: Jose Antonio Cuesta Seijo jose {- at -} KEMI {- dot -} KU {- dot -} DK
Date: 2010-04-23

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
end up mostly in the reservoir leaving onlt traces in the drop, while the
cholesterol will stay in the water. It might precipitate at first, but
should redissolve or not precipitate at all if it binds your protein.

Jose Antonio Cuesta-Seijo.

"Jerry McCully" wrote:
>
> 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 100% ethanol at a
concentration of
> 10mM. However, it is very difficult to make the dilution into 5% ethanol
either
> just in water or some buffers.
>
> Does anyone have such experience to make cholesterol solution in
normal
> buffers plus some ethanol?
>
> Thanks a lot,
>
> Jerry McCully
>
> _________________________________________________________________
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--
***************************
Jose Antonio Cuesta-Seijo

Biophysical
Chemistry Group
Department of Chemistry
University of Copenhagen
Tlf:
+45-35320261
Universitetsparken 5
DK-2100 Copenhagen,
Denmark
***************************

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: phenix error message
From: Tim Gruene tg {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2010-04-23

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From: Joachim Reichelt Joachim {- dot -} Reichelt {- at -} HELMHOLTZ-HZI {- dot -} DE
Date: 2010-04-23