Bio-Protocol (May 2017)

Detection of ASC Oligomerization by Western Blotting

  • Jérôme Lugrin,
  • Fabio Martinon

DOI
https://doi.org/10.21769/bioprotoc.2292
Journal volume & issue
Vol. 7, no. 10

Abstract

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The apoptosis-associated speck-like protein with a caspase-recruitment domain (ASC) adaptor protein bridges inflammasome sensors and caspase-1. Upon inflammasome activation, ASC nucleates in a prion-like manner into a large and single platform responsible for the recruitment and the activation of caspase-1. Active caspase-1 will in turn promote the proteolytic maturation of the pro-inflammatory cytokine IL-1β. ASC oligomerization is direct evidence for inflammasome activation and its detection allows a read-out independent of caspase-1 and IL-1β. This protocol describes how to detect the oligomerization of ASC by Western blot.