Methods to detect NF-κB acetylation and methylation

Jinjing Chen, Lin Feng Chen

Research output: Contribution to journalArticlepeer-review


Posttranslational modifications of NF-κB, including acetylation and methylation, have emerged as an important regulatory mechanism for determining the duration and strength of NF-κB nuclear activity as well as its transcriptional output. Within the seven NF-κB family proteins, the RelA subunit of NF-κB is the most studied for its regulation by lysine acetylation and methylation. Acetylation or methylation at different lysine residues modulates distinct functions of NF-κB, including DNA-binding and transcription activity, protein stability, and its interaction with NF-κB modulators. Here, we describe the experimental methods to monitor the in vitro and in vivo acetylated or methylated forms of NF-κB. These methods include radiolabeling the acetyl or methyl groups and immunoblotting with pan- or site-specific acetyl- or methyl-lysine antibodies. Radiolabeling is useful in the initial validation of the modifications. Immunoblotting with antibodies provides a rapid and powerful approach to detect and analyze the functions of these modifications in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)395-409
Number of pages15
JournalMethods in Molecular Biology
StatePublished - 2015


  • Acetylation
  • Methylation
  • NF-κB
  • Posttranslational modification
  • RelA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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