Schatzlmaier Philipp, Supper Verena, Göschl Lisa, Zwirzitz Alexander, Eckerstorfer Paul, Ellmeier Wilfried, Huppa Johannes B, Stockinger Hannes
Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria.
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.
Sci Signal. 2015 Sep 22;8(395):rs11. doi: 10.1126/scisignal.aac5584.
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.
脂筏是一类独特的高度动态的细胞膜微区，对细胞稳态、分化和信号传导至关重要。然而，当处理稀有细胞、发育上异质的细胞群体或仅与脂筏弱结合的分子时，对它们进行定量检测具有挑战性。我们提出了一种快速生化方法，该方法基于在通过非离子去污剂离心进行部分裂解时，脂筏成分与细胞核结合。我们的方案所需起始材料少且操作简便，能够以单细胞分辨率对驻留在脂筏上的蛋白质进行多维流式细胞术定量分析，从而评估复杂细胞群体中少数细胞的膜成分，以及细胞信号传导、分化或基因突变所导致的膜成分动态变化。
