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Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex
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Chapters
- 00:05Introduction
- 00:44Preparing Cells and CD95L Stimulation
- 01:11Cell Harvest and Lysis
- 02:29Immunoprecipitation
- 03:28Western Blot
- 04:21Western Blot Detection
- 05:05Analysis of CD95 DISC Formation in HeLa-CD95 cells, c-FLIPL-overexpressing HeLa-CD95 cells, and Primary T Cells
- 06:25Conclusion
Here, an experimental workflow is presented that enables the detection of caspase-8 processing directly at the death-inducing signaling complex (DISC) and determines the composition of this complex. This methodology has broad applications, from unraveling the molecular mechanisms of cell death pathways to the dynamic modeling of apoptosis networks.
