title: Rapid multiplex analysis of lipid raft components with single-cell resolution.
abstract: Lipid rafts, a distinct class of highly dynamic cell membrane microdomains, are integral to cell homeostasis, differentiation, and signaling. However, their quantitative examination is challenging when working with rare cells, developmentally heterogeneous cell populations, or molecules that only associate weakly with lipid rafts. We present a fast biochemical method, which is based on lipid raft components associating with the nucleus upon partial lysis during centrifugation through nonionic detergent. Requiring little starting material or effort, our protocol enabled the multidimensional flow cytometric quantitation of raft-resident proteins with single-cell resolution, thereby assessing the membrane components from a few cells in complex cell populations, as well as their dynamics resulting from cell signaling, differentiation, or genetic mutation.
authors.affiliation: Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria.
authors.forename: Philipp
authors.initials: P
authors.lastname: Schatzlmaier
authors.affiliation: Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria.
authors.forename: Verena
authors.initials: V
authors.lastname: Supper
authors.affiliation: Institute of Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria. Division of Rheumatology, Department of Internal Medicine III, Medical University of Vienna, Währinger Gürtel 18-20, A-1090 Vienna, Austria.
authors.forename: Lisa
authors.initials: L
authors.lastname: Göschl
authors.affiliation: Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria.
authors.forename: Alexander
authors.initials: A
authors.lastname: Zwirzitz
authors.affiliation: Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria.
authors.forename: Paul
authors.initials: P
authors.lastname: Eckerstorfer
authors.affiliation: Institute of Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria.
authors.forename: Wilfried
authors.initials: W
authors.lastname: Ellmeier
authors.affiliation: Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria. johannes.huppa@meduniwien.ac.at hannes.stockinger@meduniwien.ac.at.
authors.forename: Johannes B
authors.initials: JB
authors.lastname: Huppa
authors.affiliation: Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria. johannes.huppa@meduniwien.ac.at hannes.stockinger@meduniwien.ac.at.
authors.forename: Hannes
authors.initials: H
authors.lastname: Stockinger
mesh_terms: D002454:Cell Differentiation; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D019169:Jurkat Cells; D021962:Membrane Microdomains / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D015398:Signal Transduction
chemical_list: D008565:Membrane Proteins