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Biochemical and microscopic analysis of inflammasome complex formation
Stockholm University, Faculty of Science, Department of Molecular Biosciences, The Wenner-Gren Institute. Umeå University, Sweden.ORCID iD: 0000-0002-1269-8288
Number of Authors: 32019 (English)In: DNA Sensors and Inflammasomes / [ed] Jungsan Sohn, Elsevier, 2019, Vol. 625, p. 287-298Chapter in book (Refereed)
Abstract [en]

Inflammasomes are multiprotein signaling platforms responsible for the maturation of pro-IL-113 and pro-IL-18 as well as the induction of an inflammatory cell death termed pyroptosis. Most inflammasomes consist of an upstream sensor, in most cases an adaptor protein (ASC) and inflammatory caspases such as caspase-1. Upon activation, sensor proteins oligomerize with adaptor proteins, forming large complexes called specks. These complexes can be stabilized and detected by Western blotting or fluorescence microscopy providing a direct evidence of inflammasome activation. Here we describe protocols for two complementary methods for detecting inflammasome complexes: (1) biochemical isolation and detection of ASC oligomers by Western blot analysis and (2) microscopic visualization of active caspase-1-ASC complexes. These protocols have successfully been applied in our recent study to unveil new regulatory mechanisms for different inflammasomes including the DNA sensor AIM2 (Erttmann et al., 2016).

Place, publisher, year, edition, pages
Elsevier, 2019. Vol. 625, p. 287-298
Series
Methods in Enzymology, ISSN 0076-6879, E-ISSN 1557-7988 ; 625
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:su:diva-175913DOI: 10.1016/bs.mie.2019.05.014ISI: 000488782900018PubMedID: 31455532ISBN: 978-0-12-818359-5 (print)ISBN: 978-0-12-818360-1 (electronic)OAI: oai:DiVA.org:su-175913DiVA, id: diva2:1372209
Available from: 2019-11-22 Created: 2019-11-22 Last updated: 2019-12-04Bibliographically approved

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Gekara, Nelson O.
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CiteExportLink to record
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Citation style
  • apa
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Output format
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