Updated publication reference for PubMed record(s): 32027064. Protein AMPylation is a prevalent posttranslational modification in human cells involved in the regulation of unfolded protein response and neural development. Here we present a novel pro-N6azA probe suitable for in vivo imaging and chemical proteomics profilling of AMPylated proteins. The pro-N6azA probe provides stable fluorescent labelling in living cells accesible by straightforward strain-promoted azide-alkyne click reaction. Application of this probe may contribute significantly to the analysis of protein AMPylation during various metabolic processes including neurodevelopment and unfolded protein response.