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. |
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