Isolation and analysis of components of CD95 (APO-1/Fas) death-inducing signaling complex

Carsten Scaffidi; Peter H. Krammer; Marcus E. Peter

doi:10.1006/meth.1999.0742

Isolation and analysis of components of CD95 (APO-1/Fas) death-inducing signaling complex

Carsten Scaffidi, Peter H. Krammer, Marcus E. Peter^*

^*Corresponding author for this work

Medicine, Hematology Oncology Division

Research output: Contribution to journal › Article › peer-review

44 Scopus citations

Abstract

CD95 (APO-1/Fas) is an apoptosis-inducing receptor belonging to the tumor necrosis factor receptor superfamily. Multimerization of CD95 leads to instant recruitment of the signaling molecules FADD and caspase-8 to the activated receptor forming the death-inducing signaling complex (DISC). DISC formation is the first essential step of CD95 signaling and results in activation of caspase-8 starting a signaling cascade that leads to apoptosis. Here we describe a method for analyzing the CD95 DISC. The method is based on coimmunoprecipitation of the signaling molecules with the activated CD95 receptor followed by Western blot detection of associated molecules. Therefore, this method can analyze the very first signaling events during CD95-mediated apoptosis.

Original language	English (US)
Pages (from-to)	287-291
Number of pages	5
Journal	Methods: A Companion to Methods in Enzymology
Volume	17
Issue number	4
DOIs	https://doi.org/10.1006/meth.1999.0742
State	Published - Apr 1999

Funding

This work was supported by grants from the Deutsche For-schungsgemeinschaft, the Bundesministerium für Forschung und Technologie, and the Tumor Center Heidelberg/ Mannheim.

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
Molecular Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1006/meth.1999.0742

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title = "Isolation and analysis of components of CD95 (APO-1/Fas) death-inducing signaling complex",

abstract = "CD95 (APO-1/Fas) is an apoptosis-inducing receptor belonging to the tumor necrosis factor receptor superfamily. Multimerization of CD95 leads to instant recruitment of the signaling molecules FADD and caspase-8 to the activated receptor forming the death-inducing signaling complex (DISC). DISC formation is the first essential step of CD95 signaling and results in activation of caspase-8 starting a signaling cascade that leads to apoptosis. Here we describe a method for analyzing the CD95 DISC. The method is based on coimmunoprecipitation of the signaling molecules with the activated CD95 receptor followed by Western blot detection of associated molecules. Therefore, this method can analyze the very first signaling events during CD95-mediated apoptosis.",

author = "Carsten Scaffidi and Krammer, {Peter H.} and Peter, {Marcus E.}",

note = "Funding Information: This work was supported by grants from the Deutsche For-schungsgemeinschaft, the Bundesministerium f{\"u}r Forschung und Technologie, and the Tumor Center Heidelberg/ Mannheim.",

year = "1999",

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doi = "10.1006/meth.1999.0742",

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AU - Scaffidi, Carsten

AU - Krammer, Peter H.

AU - Peter, Marcus E.

N1 - Funding Information: This work was supported by grants from the Deutsche For-schungsgemeinschaft, the Bundesministerium für Forschung und Technologie, and the Tumor Center Heidelberg/ Mannheim.

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AB - CD95 (APO-1/Fas) is an apoptosis-inducing receptor belonging to the tumor necrosis factor receptor superfamily. Multimerization of CD95 leads to instant recruitment of the signaling molecules FADD and caspase-8 to the activated receptor forming the death-inducing signaling complex (DISC). DISC formation is the first essential step of CD95 signaling and results in activation of caspase-8 starting a signaling cascade that leads to apoptosis. Here we describe a method for analyzing the CD95 DISC. The method is based on coimmunoprecipitation of the signaling molecules with the activated CD95 receptor followed by Western blot detection of associated molecules. Therefore, this method can analyze the very first signaling events during CD95-mediated apoptosis.

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Isolation and analysis of components of CD95 (APO-1/Fas) death-inducing signaling complex

Abstract

Funding

ASJC Scopus subject areas

UN SDGs

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