| Chapter title |
Immunoprecipitation of Death Inducing Signaling Complex by Caspase-8
|
|---|---|
| Chapter number | 3 |
| Book title |
CD95
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6780-3_3 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6778-0, 978-1-4939-6780-3
|
| Authors |
Andrei Alexandru Constentinescu, Aymeric Morlé, Olivier Micheau, Andrei Alexandru Constantinescu |
| Editors |
Patrick Legembre |
| 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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