4.11. Calcein Leakage Assay

Peptide-induced membrane permeability was monitored be measuring calcein dye emission in calcein-loaded LUVs [49]. We prepared the lipid mixture, and PC:CH (2:1, w/w) and PC:CH:SM liposomes (1:1:1, w/w) were prepared with 70 mM calcein buffer. A Sephadex G-50 column was used to separate calcein-loaded liposomes from free calcein dye. The LUVs (10 μM) were mixed with various concentrations of the peptide on a 96-well black plate. The fluorescence intensity was measured at excitation and emission wavelengths of 480 and 520 nm, respectively.