Phos-tag-based non-radioactive protocols for monitoring Arabidopsis kinase activities in vitro

Abstract

Kinases are indispensable signaling components. Radioactive-based phosphorylation assays are widely used but require specific protective equipment and safety trainings. Here, we present a Phos-tag-based non-radioactive kinase assay to study Arabidopsis kinase activities. We expressed and purified both kinase and substrate proteins from E. coli cells and then used the Phos-tag technology to detect the kinase activities under either different temperatures or chemical treatments. This non-radioactive approach is environmentally friendly and applicable to other kinases and organisms. For complete details on the use and execution of this protocol, please refer to Lin et al. (2022).

Keywords: Model organisms; Plant sciences; Protein biochemistry; Signal transduction.