Supported Lipid Bilayer Technology for the Study of Cellular Interfaces

Overview

Journal Curr Protoc Cell Biol

Specialty Cell Biology

Date 2015 Sep 3

PMID 26331983

Citations 11

Authors

Travis J Crites

Michael Maddox

Kartika Padhan

James Muller

Calvin Eigsti

Rajat Varma

Affiliations

Soon will be listed here.

Abstract

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.

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