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.
The original version of this chapter was revised. The erratum to this chapter is available at: DOI 10.1007/978-1-4939-6780-3_22
An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-4939-6780-3_22
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Constantinescu, A.A., Morlé, A., Micheau, O. (2017). Immunoprecipitation of Death Inducing Signaling Complex by Caspase-8. In: Legembre, P. (eds) CD95. Methods in Molecular Biology, vol 1557. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6780-3_3
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DOI: https://doi.org/10.1007/978-1-4939-6780-3_3
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