Protocol for isolation of microbiota-derived membrane vesicles from mouse blood and colon.
| Autor: | Erttmann SF; 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. Electronic address: s.erttmann@ikmb.uni-kiel.de., Gekara NO; 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. Electronic address: nelson.ongondo.gekara@uniklinik-freiburg.de. |
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| Jazyk: | angličtina |
| Zdroj: | STAR protocols [STAR Protoc] 2023 Mar 17; Vol. 4 (1), pp. 102046. Date of Electronic Publication: 2023 Jan 19. |
| DOI: | 10.1016/j.xpro.2023.102046 |
| Abstrakt: | 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). 1 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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