Unit

UNIT 18.13 Supported Planar Bilayers for Study of the Immunological Synapse

  1. Michael L. Dustin,
  2. Toby Starr,
  3. Rajat Varma,
  4. V. Kaye Thomas

Published Online: 1 FEB 2007

DOI: 10.1002/0471142735.im1813s76

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Dustin, M. L., Starr, T., Varma, R. and Thomas, V. K. 2007. Supported Planar Bilayers for Study of the Immunological Synapse. Current Protocols in Immunology. 76:18.13:18.13.1–18.13.35.

Author Information

  1. Skirball Institute of Biomolecular Medicine, New York, New York

Publication History

  1. Published Online: 1 FEB 2007
  2. Published Print: FEB 2007

Abstract

Supported planar bilayers have been used in immunology research for over 25 years, including in the initial demonstrations of MHC-peptide complex functional activity and adhesion molecule activity. More recent modifications of the method have been used to measure two-dimensional affinities and to study the formation of the immunological synapse. This unit covers the incorporation of glycolipid-anchored membrane proteins, 6-histidine-tagged soluble proteins, and monobiotinylated soluble proteins into supported planar bilayers. Reagents developed for the MHC-peptide tetramer staining method (unit 17.3) can readily be adapted to presentation on planar bilayers. The unique advantage of this approach is that the proteins presented on the surface of the supported bilayer are laterally mobile. This provides a more physiological presentation of cell-surface molecules and supports visualization of protein rearrangement on the bilayer by live cells.

Keywords:

  • microscopy;
  • receptors;
  • affinity;
  • fluorescence;
  • signaling;
  • activation