Method to Measure Ubiquitination of NLRs

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 N...

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Bibliographic Details
Published in:	Methods in molecular biology (Clifton, N.J.) Vol. 1417; p. 223
Main Authors:	Palazón-Riquelme, Pablo, López-Castejón, Gloria
Format:	Journal Article
Language:	English
Published:	United States 01-01-2016
Subjects:	Blotting, Western HEK293 Cells Humans Inflammasomes - metabolism NLR Family, Pyrin Domain-Containing 3 Protein - metabolism Ubiquitination NLRP3 Ubiquitination Inflammasome Immunoprecipitation TUBE
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Summary:	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.
ISSN:	1940-6029
DOI:	10.1007/978-1-4939-3566-6_16