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Updated: Mar 9, 2026

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex
Published on: August 2, 2021
Immunoprecipitation of Death Inducing Signaling Complex by Caspase-8
Andrei Alexandru Constantinescu1,2, Aymeric Morlé1,2, Olivier Micheau3,4
1INSERM, UMR866, "Equipe labellisée Ligue contre le Cancer" and Laboratoire d'Excellence LipSTIC, Dijon, 21079, France.
Abstract:
Analysis of CD95/Fas complexes by immunoprecipitation has long relied on the monoclonal antibody APO1 or tagged recombinant Fas ligand. Immunoprecipitation is an elegant and efficient procedure to investigate endogenous protein interactions or complexes. Provided that the targeted complex is soluble in mild detergent these complexes can be recovered using protein A/G-coupled Sepharose beads and further analyzed after denaturation and electrophoretic separation by western blotting or mass spectrometry. Herein, we describe in detail the method used in our laboratory to immunoprecipitate and analyze by immunoblot complexes containing caspase-8, using a commercial antibody directed against caspase-8.
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