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. 2021:2318:69-85.
doi: 10.1007/978-1-0716-1476-1_5.

Detection of Post-translational Modifications on MYC

Colin J Daniel  1 , Xiao-Xin Sun  1 , Yingxiao Chen  1 , Xiaoli Zhang  1 , Mu-Shui Dai  2   3 , Rosalie C Sears  4   5
Affiliations

Detection of Post-translational Modifications on MYC

Colin J Daniel et al. Methods Mol Biol. 2021.

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.

Keywords: Immunofluorescence; Immunoprecipitation; Phosphorylation; SUMOylation; Ubiquitination; c-Myc.

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