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).
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Acknowledgments
This work was supported by the Austrian FWF grants P25691 to S.S. and W901-B12 (DK Molecular Enzymology) to P.K., F.Z., and S.S. as well as BioTechMed Graz (Flagship project “Secretome”).
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Kohl, P., Zingl, F.G., Eichmann, T.O., Schild, S. (2018). Isolation of Outer Membrane Vesicles Including Their Quantitative and Qualitative Analyses. In: Sikora, A. (eds) Vibrio Cholerae. Methods in Molecular Biology, vol 1839. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8685-9_11
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DOI: https://doi.org/10.1007/978-1-4939-8685-9_11
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