Detection of RIPK1 in the FADD-containing death inducing signaling complex (DISC) during necroptosis

Rosalind L. Ang, Adrian T. Ting

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

FAS-associated protein with death domain (FADD) is a signaling molecule required by members of the TNF receptor superfamily (TNFRSF) such as FAS and TNFR1 to induce apoptosis. FADD is a small adapter molecule that functions as a scaffold to recruit procaspase-8 and other regulators. The FADD-containing signaling complex that initiates the apoptotic cascade has been termed the death inducing signaling complex (DISC). In the absence of FADD, death receptors cannot induce apoptosis and in appropriate cell types, these death receptors then induce necroptosis. Necroptosis can also be induced by death receptors in FADD-sufficient cells when caspase-8 is inhibited, usually accomplished by the addition of caspase inhibitors. Under such necroptotic conditions, the immunoprecipitation of FADD to isolate the DISC can be utilized to examine components of this complex. Here, we describe the immunoprecipitation of FADD and subsequent western-blotting to identify RIPK1 in this complex during necroptosis.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages101-107
Number of pages7
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1857
ISSN (Print)1064-3745

Keywords

  • Coimmunoprecipitation
  • Death inducing signaling complex (DISC)
  • FAS-associated protein with death domain (FADD)
  • Receptor-interacting protein kinase 1 (RIPK1)
  • Western blotting

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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