Supported Lipid Bilayer Technology for the Study of Cellular Interfaces
Glass‐supported lipid bilayers presenting freely diffusing proteins have served as a powerful tool for studying cell‐cell interfaces, in particular, T cell–antigen presenting cell (APC) interactions, using optical microscopy. Here we expand upon existing protocols and describe the preparation of lip...
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| Veröffentlicht in: | Current protocols in cell biology 2015-09, Vol.68 (1), p.24.5.1-24.5.31 |
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| container_end_page | 24.5.31 |
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| container_issue | 1 |
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| container_title | Current protocols in cell biology |
| container_volume | 68 |
| creator | Crites, Travis J. Maddox, Michael Padhan, Kartika Muller, James Eigsti, Calvin Varma, Rajat |
| description | Glass‐supported lipid bilayers presenting freely diffusing proteins have served as a powerful tool for studying cell‐cell interfaces, in particular, T cell–antigen presenting cell (APC) interactions, using optical microscopy. Here we expand upon existing protocols and describe the preparation of liposomes by an extrusion method, and describe how this system can be used to study immune synapse formation by Jurkat cells. We also present a method for forming such lipid bilayers on silica beads for the study of signaling responses by population methods, such as western blotting, flow cytometry, and gene‐expression analysis. Finally, we describe how to design and prepare transmembrane‐anchored protein‐laden liposomes, following expression in suspension CHO (CHOs) cells, a mammalian expression system alternative to insect and bacterial cell lines, which do not produce mammalian glycosylation patterns. Such transmembrane‐anchored proteins may have many novel applications in cell biology and immunology. © 2015 by John Wiley & Sons, Inc. |
| doi_str_mv | 10.1002/0471143030.cb2405s68 |
| format | Article |
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| identifier | ISSN: 1934-2500 |
| ispartof | Current protocols in cell biology, 2015-09, Vol.68 (1), p.24.5.1-24.5.31 |
| issn | 1934-2500 1934-2616 |
| language | eng |
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| source | MEDLINE; Alma/SFX Local Collection |
| subjects | Animals CHO Cells CHOs cells Cricetulus diffusion Lipid Bilayers liposome extrusion Liposomes - chemistry Membrane Proteins - metabolism protein engineering Protein Engineering - methods Recombinant Proteins - metabolism recombinant transmembrane proteins signal transduction supported lipid bilayers suspension CHO cells synapse |
| title | Supported Lipid Bilayer Technology for the Study of Cellular Interfaces |
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