Detection of post-translational modifications on MYC

Colin J. Daniel, Xiao Xin Sun, Yingxiao Chen, Xiaoli Zhang, Mu Shui Dai, Rosalie C. Sears

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Detection of post-translational modifications in c-Myc is an invaluable tool in assessing Myc status, particularly in cancer. However, it can be challenging to detect these modifications. The evaluation of phosphorylation status of c-Myc can also be challenging with the current commercially available phosphorylation sensitive antibodies. Here, we describe protocols for the immunoprecipitation of endogenous c-Myc to probe for phosphorylation status, as well as the detection of ubiquitination and SUMOylation on c-Myc. We will also discuss the challenges of detecting phosphorylated c-Myc in formalin-fixed paraffin-embedded tissues by immunofluorescence and describe a protocol using a new rat monoclonal antibody we have generated suitable for this purpose.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages69-85
Number of pages17
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2318
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Immunofluorescence
  • Immunoprecipitation
  • Phosphorylation
  • SUMOylation
  • Ubiquitination
  • c-Myc

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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