3 Methods
3.1 Small
Unilamellar Liposome
Preparation
1. An x mg/mL lipid small unilamellar vesicle (SUV) stock solution is prepared by dissolving x mg of lipid in chloroform
(or other solvent of choice). Dry the lipid onto the walls of a
round-bottom flask (see Notes 1 and 2). The analyte(s) may be
incorporated at this stage as powder or concentrated solution
in a volatile solvent or added later (step 5).
2. The film is then resuspended by adding 1 mL of the required
buffer (including salts, see Note 3) to give a turbid suspension
at the desired stock concentration.
3. Five cycles of freeze/thawing are then implemented immediately after one another. A convenient freezing method is to use
Fig. 5 Examples of lipids (available from Avanti (Alabaster, AL, USA)) that can form bilayers. DMPC, DPPC,
POPC, and POPS denote, respectively: 1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, and 1-palmitoyl-2-oleoyl-snglycero-3-phospho-L-serine
Flow LD of Protein–Membrane Systems
455
3.1 Small
Unilamellar Liposome
Preparation
1. An x mg/mL lipid small unilamellar vesicle (SUV) stock solution is prepared by dissolving x mg of lipid in chloroform
(or other solvent of choice). Dry the lipid onto the walls of a
round-bottom flask (see Notes 1 and 2). The analyte(s) may be
incorporated at this stage as powder or concentrated solution
in a volatile solvent or added later (step 5).
2. The film is then resuspended by adding 1 mL of the required
buffer (including salts, see Note 3) to give a turbid suspension
at the desired stock concentration.
3. Five cycles of freeze/thawing are then implemented immediately after one another. A convenient freezing method is to use
Fig. 5 Examples of lipids (available from Avanti (Alabaster, AL, USA)) that can form bilayers. DMPC, DPPC,
POPC, and POPS denote, respectively: 1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, and 1-palmitoyl-2-oleoyl-snglycero-3-phospho-L-serine
Flow LD of Protein–Membrane Systems
455