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Biochemical and microscopic analysis of inflammasome complex formation
Stockholms universitet, Naturvetenskapliga fakulteten, Institutionen för molekylär biovetenskap, Wenner-Grens institut. Umeå University, Sweden.ORCID-id: 0000-0002-1269-8288
Rekke forfattare: 32019 (engelsk)Inngår i: DNA Sensors and Inflammasomes / [ed] Jungsan Sohn, Elsevier, 2019, Vol. 625, s. 287-298Kapittel i bok, del av antologi (Fagfellevurdert)
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).

sted, utgiver, år, opplag, sider
Elsevier, 2019. Vol. 625, s. 287-298
Serie
Methods in Enzymology, ISSN 0076-6879, E-ISSN 1557-7988 ; 625
HSV kategori
Identifikatorer
URN: urn:nbn:se:su:diva-175913DOI: 10.1016/bs.mie.2019.05.014ISI: 000488782900018PubMedID: 31455532ISBN: 978-0-12-818359-5 (tryckt)ISBN: 978-0-12-818360-1 (digital)OAI: oai:DiVA.org:su-175913DiVA, id: diva2:1372209
Tilgjengelig fra: 2019-11-22 Laget: 2019-11-22 Sist oppdatert: 2019-12-04bibliografisk kontrollert

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