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Quantification of Protein Kinase Activities by LC-MS

  • Maria P. Alcolea
  • Pedro R. Cutillas
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 658)

Abstract

Measuring the enzymatic activity of protein kinases in cell and tissue extracts represents a difficult task owing to the complex regulation and dynamics of such enzymes. Here we describe a sensitive and specific approach for the quantitative analysis of PI3K-dependent protein kinase activity based on the mass spectrometry measurement of reaction products. The principle of this method can be applied to develop other kinase assays and thus should contribute to the understanding of processes controlled by protein kinases. Because of the enhanced sensitivity of this technique, it may be applied to the multiplex measurement of pathway activities when sample amounts are limiting.

Key words

PI3K signalling pathway AKT/PKB kinase enzymatic assay strong cation exchange (SCX) kinase activity quantification mass spectrometry cancer 

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Maria P. Alcolea
    • 1
  • Pedro R. Cutillas
    • 1
  1. 1.Analytical Signalling Group, Centre for Cell SignallingInstitute of Cancer, Bart’s and the London School of Medicine, Queen Mary University of LondonLondonUK

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