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 poly-ubiquitination of NRLP3 using two different approaches: (i) detection with a ubiquitin antibody or (ii) using TUBEs (Tandem Ubiquitin Binding entities). This approach can be used to detect ubiquitination of other NLRs or other proteins.
Keywords: Immunoprecipitation; Inflammasome; NLRP3; TUBE; Ubiquitination.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.