(18) For BSA-treated experiments, the vesicles must be incubated with BSA (1 mol% with respect to the lipid concentration) for 20 min before use. Due to the destabilisation of the membrane from the adsorption of BSA and the sequestration of free fatty acids, consideration must be given to the amount of lipids that should be treated since leaking will occur overnight. Therefore, we recommend that the volume of lipids to be treated does not exceed the number of experiments that can be conducted in a single day. If more BSA experiments need to be performed, treat more of the stored lipid stock.