Department of Molecular Biology, Umeå Centre for Microbial Research (UCMR), Umeå University, 90187 Umeå, Sweden; Infection Oncology Unit, Institute of Clinical Molecular Biology, Christian-Albrechts University of Kiel, UKSH, Schwanenweg 20, 24105 Kiel, Germany.
Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, 106 91 Stockholm, Sweden; Faculty of Medicine, Institute of Medical Microbiology and Hygiene, Medical Center, University of Freiburg, Hermann-Herder-Str. 11, 79104 Freiburg, Germany.
STAR Protoc. 2023 Mar 17;4(1):102046. doi: 10.1016/j.xpro.2023.102046. Epub 2023 Jan 19.
Bacterial membrane vesicles have emerged as gadgets allowing remote communication between the microbiota and distal host organs. Here we describe a protocol for enriching vesicles from serum and colon that could widely be adapted for other tissues. We detail pre-clearing of serum or colon fluids using 0.2-μm syringe filters and their concentration by centrifugal filter devices. We also describe vesicle isolation with qEV size exclusion columns and finally the concentration of isolated vesicle fractions for downstream analyses. For complete details on the use and execution of this protocol, please refer to Erttmann et al. (2022)..
细菌膜泡已经成为一种工具，允许微生物群和远端宿主器官之间进行远程通信。在这里，我们描述了一种从血清和结肠中富集囊泡的方案，该方案可以广泛适用于其他组织。我们详细介绍了使用 0.2μm 注射器过滤器对血清或结肠液进行预澄清，以及使用离心过滤装置进行浓缩的方法。我们还描述了使用 qEV 排阻柱分离囊泡，以及最后对分离的囊泡级分进行浓缩用于下游分析的方法。有关该方案使用和实施的完整详细信息，请参阅 Erttmann 等人。（2022）。
