Visualization of fas-mediated death-inducing signaling complex formation by immunoprecipitation

Methods Mol Biol. 2013:979:43-9. doi: 10.1007/978-1-62703-290-2_5.

Abstract

The prototypical death receptor Fas (also known as CD95 or Apo-1) plays an essential role in the maintenance of lymphocyte homeostasis. Propagation of cell death through Fas relies on the formation of a multiprotein complex at the receptor level known as the death-inducing signaling complex (DISC). Here, we describe an immunoprecipitation-based protocol to study DISC assembly in activated human T lymphocytes. This procedure is a useful tool to visualize proteins associated with Fas.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Collodion / chemistry
  • Death Domain Receptor Signaling Adaptor Proteins / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Immunoblotting
  • Immunoprecipitation / methods*
  • Lymphocyte Activation
  • Membranes, Artificial
  • Molecular Imaging / methods*
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • fas Receptor / metabolism*

Substances

  • Death Domain Receptor Signaling Adaptor Proteins
  • Membranes, Artificial
  • fas Receptor
  • Collodion