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R.E.W. Hancock Laboratory: Methods Liposome Preparation

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:
  1. Dry 0.5 mmole of dioleoyl phosphatidyl choline under nitrogen in a disposable glass tube.
  2. Evacuate in dessicator under vacuum for 30 minutes.
  3. Add buffer / dH20 to required volume and scrape the sides of the glass tube to dislodge the lipid.
  4. Add protein at 1 mg/ul of lipid used.
  5. 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.