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Assessing Kinase Activity in Plants with In-Gel Kinase Assays

  • Pengcheng Wang
  • Jian-Kang ZhuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1363)

Abstract

The in-gel protein kinase assay is a powerful method to measure the protein phosphorylation activity of specific protein kinases. Any protein substrate can be embedded in polyacrylamide gels where they can be phosphorylated by protein kinases that are separated in the gel under denaturing conditions and then renatured. The kinase activity can be visualized in situ in the gels by autoradiography. This method has been used to compare the activities of protein kinases in parallel samples or to identify their potential substrates. Here, we describe in detail an in-gel kinase assay to measure the activity of some protein kinases in plants.

Key words

Protein kinase Substrate Autoradiography MAPK CDPK SnRK2 Casein kinase 

Notes

Acknowledgements

This work was supported by NIH grant R01GM059138 to J-KZ.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Horticulture and Landscape ArchitecturePurdue UniversityWest LafayetteUSA
  2. 2.Shanghai Center for Plant Stress Biology, Chinese Academy of SciencesShanghai Institutes for Biological SciencesShanghaiChina

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