In article (Dans l'article) <3426C57F.787B at ibex.ca>, reckteac at ibex.ca
wrote (écrivait) :
> I am looking for methods to delipidate protein solutions, or protein &
> cell-suspensions after cell-disruption. These methods should be scalable
> for use in a large-scale protein purification process.
I am successfully using chloroform:methanol (1:2) with either membrane
preparations of detergent extracts. For both, the CHCl3:MeOH volume is
about 4 times greater than the volume of the pellet/extract. After
centrifugation, you would then get your proteins as a precipitate
saturated with CHCl3:MeOH, which are easily evaporated under nitrogen. The
dry pellet can be resuspended with any appropriate method (sonication,
potter-elvejhem, +/- detergent).
This works fine with my lipoprotein. If you need co-factors for your
protein, be sure to add them during the final resolubilization process.
good luck !
_ _ _
\/ \/
{O} Olivier Nosjean
|_| Laboratoire de Physico-Chime Biologique
||| Univ. C. Bernard - Lyon 1
| France
