Liposomes for ion transport - questions, questions, questions

Mike Holloway holloway at cris.com
Mon Jun 24 12:22:32 EST 1996


I need to make unilamellar vesicles in order to imbed a peptide and test its
ability to act as an ion channel.  I'm measuring Ca transport by loading
fluoro-3 in the vesicles.  So far I've tried 9:1 PS and PE by the method of
Huber, et al., FEBS 317, 131, and substituting 9:1 PC and PG in the same
method.  Both vesicle types are leaky, reaching maximal fluorescence within
minutes when Ca is added.  

I was wondering if anyone could share any helpful experience.  I've been
drying down the lipids on any old beaker on the shelf.  Does the glassware
have to be specially prepared?  Are there special precautions that have to be
taken to avoid detergent contamination, or something else that could make the
vesicles leaky?  Is it necessary to carefully buffer the Ca with EGTA?  

Thanks,
Mike Holloway
mike.holloway at stjude.org



More information about the Methods mailing list