title: Method to Measure Ubiquitination of NLRs.
abstract: Posttranslational modifications are crucial in determining the functions of proteins in the cell. Modification of the NLRP3 inflammasome by the ubiquitin system has recently emerged as a new level of regulation of the inflammasome complex. Here, we describe a method to detect polyubiquitination of NRLP3 using two different approaches: (1) detection with an ubiquin antibody or (2) using TUBE (Tandem Ubiquitin Binding entities). This approach can be used to detect ubiquitination of other NLR or other components of the inflammasome.
authors.affiliation: Manchester Collaborative Centre of Inflammation Research, The University of Manchester, Manchester, M13 9NT, UK.
authors.forename: Pablo
authors.initials: P
authors.lastname: Palazón-Riquelme
authors.affiliation: Manchester Collaborative Centre of Inflammation Research, The University of Manchester, Manchester, M13 9NT, UK. Gloria.Lopez-Castejon@manchester.ac.uk.
authors.forename: Gloria
authors.initials: G
authors.lastname: López-Castejón
mesh_terms: D015153:Blotting, Western; D057809:HEK293 Cells; D006801:Humans; D058847:Inflammasomes / Q000378:metabolism; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D058847:Inflammasomes; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human
keywords: Immunoprecipitation; Inflammasome; NLRP3; TUBE; Ubiquitination