Biotin lc bsa

GUVs were prepared under physiological salt conditions (20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid [HEPES], pH 7.5, 150 mM KCl) using a modified electroformation protocol and laboratory-built platinum wire electrode chambers as described previously (Montes et al., 2007 (link)). The lipid mixture contained, in addition to DOPC (∼75 mol%), 5 mol% PIP₂, 20 mol% 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (DOPS), 0.1 mol% RhPE for fluorescence imaging, and 0.5 mol% biotin-PE for GUV adhesion to streptavidin (Sigma-Aldrich, St. Louis, MO)–coupled, biotin-LC-BSA (Thermo Scientific, Rockford, IL)–coated LabTek II chambered cover slides (Nalge Nunc International, Rochester, NY) prepared according to published protocols (Yildiz et al., 2003 (link)). Confocal images were captured using a 60× oil-immersion objective and an Olympus FV1000 IX81 confocal microscope (Olympus USA, Melville, NY).