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Caspases01:24

Caspases

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Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
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Related Experiment Video

Updated: Mar 24, 2026

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex
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Ripoptosome Analysis by Caspase-8 Coimmunoprecipitation.

Maria Feoktistova1, Peter Geserick1, Martin Leverkus2

  • 1Section of Molecular Dermatology, Department of Dermatology, Venereology, and Allergology, Medical Faculty Mannheim, University Heidelberg, Heidelberg 68167, Germany;

Cold Spring Harbor Protocols
|March 3, 2016
PubMed
Summary

Researchers developed a co-immunoprecipitation method using caspase-8 antibodies to detect the Ripoptosome complex, crucial for cell death pathways like apoptosis and necroptosis.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Cell death pathways rely on high-molecular mass protein complexes.
  • Understanding the assembly and composition of these complexes is key to elucidating their roles in apoptosis and necroptosis.

Purpose of the Study:

  • To describe a co-immunoprecipitation protocol for studying the Ripoptosome complex.
  • To determine the formation and composition of the Ripoptosome in HaCaT cells.

Main Methods:

  • Caspase-8 co-immunoprecipitation was employed in HaCaT keratinocyte cell line.
  • The protocol utilizes caspase-8 antibodies to isolate the Ripoptosome complex.

Main Results:

  • The method successfully detects the formation and composition of the Ripoptosome complex.
  • Caspase-8 is confirmed as an essential component of the Ripoptosome.

Conclusions:

  • This protocol enables the investigation of Ripoptosome formation and composition.
  • The method can be adapted for other antibodies and cell lines to study cell death signaling complexes.