The apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC) functions as the integral adaptor protein between inflammasome sensors such as NOD-like receptor protein 3 (NLRP3) and the inflammatory caspase, caspase-1. Inflammasome sensor triggering allows recruitment of ASC and the formation of long amyloid-like ASC oligomers that enable binding and proximity-induced activation of caspase-1. The detection of ASC oligomerization thus constitutes a highly specific and direct test for inflammasome complex formation and activation. Here, we describe a simplified and streamlined method for the detection of ASC oligomers via Western blotting, using the chemical crosslinking reagent disuccinimidyl suberate.
Keywords: ASC; Caspase-1; Cell death; Inflammasome; Inflammation; Innate immunity; Interleukin-1; NLRP3; Pyroptosis.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.