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A Streamlined Method for Detecting Inflammasome-Induced ASC Oligomerization Using Chemical Crosslinking

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Inflammation and Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2691))

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Abstract

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.

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Acknowledgments

This work was funded by a National Health and Medical Research Council (NHMRC) of Australia Ideas Grant (1183070) and Investigator Grant (1172929). The contents of the published materials are solely the responsibility of the individual authors and do not reflect the views of the NHMRC. This work was also supported by operational infrastructure grants through the Australian Government Independent Research Institute Infrastructure Support Scheme (9000719) and the Victorian State Government Operational Infrastructure Support, Australia.

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Correspondence to James E. Vince .

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Hughes, S.A., Vince, J.E. (2023). A Streamlined Method for Detecting Inflammasome-Induced ASC Oligomerization Using Chemical Crosslinking. In: Jenkins, B.J. (eds) Inflammation and Cancer. Methods in Molecular Biology, vol 2691. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3331-1_12

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  • DOI: https://doi.org/10.1007/978-1-0716-3331-1_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3330-4

  • Online ISBN: 978-1-0716-3331-1

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