Stock solutions (1–10 mg/mL)
of 1-palmitoyl-2-oleoyl-
sn-glycero-3-phosphocholine
(POPC), 1-palmitoyl-2-oleoyl-
sn-glycero-3-phospho-
l-serine (POPS, Avanti Polar Lipids, Alabaster, AL, USA), and
ATTO 390-1,2-dipalmitoyl-
sn-glycero-3-phosphoethanolamine
(Atto 390-DPPE, ATTO-TEC, Siegen, Germany) were prepared in chloroform.
l-α-Phosphatidylinositol-4,5-bisphosphate (PtdIns[4,5]P
2, brain porcine, Avanti Polar Lipids, Alabaster, AL, USA)
was freshly dissolved in chloroform/methanol/H
2O (10:20:8)
to a final concentration of 1 mg/mL. Lipid mixtures (0.4 mg) were
prepared in chloroform/methanol (10:1), and organic solvents were
evaporated with a nitrogen stream followed by 3 h in vacuum. The dried
lipid films were stored at 4 °C until needed.
Small unilamellar
vesicles (SUVs) were prepared by rehydrating a lipid film in spreading
buffer (50 mM KCl, 20 mM Na-citrate, 0.1 mM NaN
3, 0.1 mM
ethylenediaminetetraacetic acid (EDTA), pH 4.8),
38 (link) incubating for 30 min, subsequent vortexing (3 × 30
s at 5 min intervals), and a final sonification step for 30 min at
room temperature (cycle 4, 60%, Sonopuls HD2070, resonator cup; Bandelin,
Berlin, Germany). PtdIns[4,5]P
2 containing SUVs were used
immediately for the preparation of SLBs to avoid PtdIns[4,5]P
2 degradation.
65 (link)
Liebe N.L., Mey I., Vuong L., Shikho F., Geil B., Janshoff A, & Steinem C. (2023). Bioinspired Membrane Interfaces: Controlling Actomyosin Architecture and Contractility. ACS Applied Materials & Interfaces, 15(9), 11586-11598.