Using sequential immunoprecipitation and mass spectrometry to identify methylation of NF-κB.
Posttranslational modifications have long been known to play an essential role in the regulation of NF-κB activity. In the past few years, in addition to more traditional modifications such as phosphorylation, the p65 subunit of NF-κB has been found to be methylated at multiple sites. Here, we describe procedures for using immunoprecipitation and mass spectrometry to identify the methylation sites of p65.
Department of Pharmacology and Toxicology, Indiana University School of Medicine, 635 Barnhill Drive, Indianapolis, IN, 46202, USA, lut@iupui.edu.
Tao
T
Lu
George R
GR
Stark
D018929:Cell Culture Techniques; D002460:Cell Line; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D066298:In Vitro Techniques; D053583:Interleukin-1beta / Q000494:pharmacology; D013058:Mass Spectrometry / Q000379:methods; D008745:Methylation / Q000187:drug effects; D016328:NF-kappa B / Q000378:metabolism*; D010766:Phosphorylation; D051996:Transcription Factor RelA / Q000378:metabolism
D053583:Interleukin-1beta; D016328:NF-kappa B; D051996:Transcription Factor RelA