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Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex
JoVE 杂志
生物化学
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JoVE 杂志 生物化学
Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex
DOI:

07:17 min

August 02, 2021

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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

Summary

自动翻译

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.

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