{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T12:36:33Z","timestamp":1790944593975,"version":"4.1.0"},"reference-count":28,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2010,3,1]],"date-time":"2010-03-01T00:00:00Z","timestamp":1267401600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CP Cell Biology"],"published-print":{"date-parts":[[2010,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The environmental bacterium <jats:named-content content-type=\"genus-species\"><jats:italic>Legionella pneumophila<\/jats:italic><\/jats:named-content> naturally parasitizes free\u2010living amoebae. <jats:named-content content-type=\"genus-species\"><jats:italic>L. pneumophila<\/jats:italic><\/jats:named-content> is an opportunistic human pathogen that grows in macrophages, thus causing a life\u2010threatening pneumonia termed Legionnaires' disease. The bacteria replicate intracellularly in environmental and immune phagocytes within a unique compartment, the <jats:italic>Legionella<\/jats:italic>\u2010containing vacuole (LCV). Formation of LCVs is a complex and robust process involving &gt;150 secreted bacterial effector proteins, which are believed to subvert host cell signaling and vesicle trafficking pathways. This unit describes a simple approach to purify intact LCVs from <jats:named-content content-type=\"genus-species\"><jats:italic>Dictyostelium discoideum<\/jats:italic><\/jats:named-content> amoebae. The method comprises a two\u2010step purification protocol that includes immuno\u2010magnetic separation by means of an antibody against an effector protein specifically binding to LCVs, followed by density gradient centrifugation. The use of <jats:named-content content-type=\"genus-species\"><jats:italic>D. discoideum<\/jats:italic><\/jats:named-content> producing a fluorescent LCV marker and fluorescently labeled <jats:named-content content-type=\"genus-species\"><jats:italic>L. pneumophila<\/jats:italic><\/jats:named-content> allow tracking the enrichment of LCVs by light microscopy. <jats:italic>Curr. Protoc. Cell Biol<\/jats:italic>. 46:3.34.1\u20103.34.14. \u00a9 2010 by John Wiley &amp; Sons, Inc.<\/jats:p>","DOI":"10.1002\/0471143030.cb0334s46","type":"journal-article","created":{"date-parts":[[2010,3,16]],"date-time":"2010-03-16T18:26:15Z","timestamp":1268763975000},"source":"Crossref","is-referenced-by-count":18,"title":["Isolation of <i>Legionella<\/i>\u2010Containing Vacuoles by Immuno\u2010Magnetic Separation"],"prefix":"10.1002","volume":"46","author":[{"given":"Simon","family":"Urwyler","sequence":"first","affiliation":[{"name":"Institute of Molecular Life Sciences, University of Z\u00fcrich  Z\u00fcrich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ivo","family":"Finsel","sequence":"additional","affiliation":[{"name":"Institute of Molecular Life Sciences, University of Z\u00fcrich  Z\u00fcrich 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