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Quantifying the Kinase Activities of MST1/2

  • Niamh A. O’Driscoll
  • David MatallanasEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1893)

Abstract

The functions of the kinases MST1 and MST2 rely heavily on their ability to phosphorylate and become phosphorylated themselves. Hence, it is important to precisely measure the kinase activities of both isoforms in a reproducible manner. Here, we describe in detail the protocol for an in-gel kinase assay for the quantification of the kinase activity of MST1/2, which involves immunoprecipitation of MST1/2 and the incorporation of radiolabeled phosphate from [γ-32P]-ATP into a substrate immobilized in a polyacrylamide gel. We also include a protocol for indirect measurement of MST1/2 activation status using immunoblotting.

Key words

MST1 MST2 Kinase assay Immunoprecipitation Radiolabel 

Notes

Acknowledgments

This work was supported by Science Foundation Ireland grants 15/CDA/3495 and 16/TIDA/4064.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Systems Biology IrelandUniversity College Dublin, BelfieldDublinIreland
  2. 2.School of MedicineUniversity College Dublin, BelfieldDublinIreland

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