Liposome Preparation
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OBJECTIVE:
Method for incorporating proteins into liposomes for liposome swelling assay or as an alternative antigen presentation method.
REAGENTS:
Dioleoyl Phosphatidyl Choline
Buffer of choice / distilled water
METHODS:
Dry 0.5 mmole of dioleoyl phosphatidyl choline under nitrogen in a disposable glass tube.
Evacuate in dessicator under vacuum for 30 minutes.
Add buffer / dH20 to required volume and scrape the sides of the glass tube to dislodge the lipid.
Add protein at 1 mg/ul of lipid used.
Vortex for 30 seconds. Sonicate twice in a bath sonicator at 7 degree for 15 sec.
This makes multilamellar vesicles that become small unilamellar vesicles (SUV) with prolonged sonication time. To make large unilamellar vesicles, use the extruder.
