Isolation of outer membrane vesicles including their quantitative and qualitative analyses

Abstract

Outer membrane vesicles (OMVs) are naturally secreted from the bacterial cell surface and therefore localized in the cell-free supernatant of bacterial cultures. Here we describe methods for crude and density gradient-purified OMV isolation and protocols for control analyses for protein profiling (SDS-PAGE), detection of indicator proteins (immunoblot analysis), lipid profiling (lipid extraction and LC-MS analysis), vesicle size determination (NanoSight), rough estimation of biomass (TrayCell™), as well as quantifications of defined OMV components, e.g., proteins (Bradford) and LPS (Purpald).