Kinase activity determination of specific AMPK complexes/heterotrimers in the skeletal muscle

1 Citation (Scopus)

Abstract

Measuring the kinase activity of the 5'-AMP-activated protein kinase (AMPK) is an essential part of understanding the regulation of this metabolic master switch. The AMPK heterotrimer can exist in 12 different constellations with potentially diverse activation patterns. It is therefore important to be able to measure heterotrimer-specific activity to discriminate between these patterns. In this chapter we describe how to measure the AMPK activity of specific heterotrimeric complexes by consecutive immunoprecipitations and how the assay can be performed in a medium throughput fashion using 96-well plates.

Original languageEnglish
Title of host publicationAMPK : Methods and Protecols
EditorsDietbert Neumann, Benoit Viollet
Number of pages14
Place of PublicationNew York
PublisherHumana Press
Publication date2018
Pages215-228
Chapter14
ISBN (Print)978-1-4939-7597-6
ISBN (Electronic)978-1-4939-7598-3
DOIs
Publication statusPublished - 2018
SeriesMethods in Molecular Biology
Number1732
ISSN1064-3745

Keywords

  • Faculty of Science
  • Immunoprecipitation
  • AMPK activity
  • Skeletal muscle
  • Isoform-specific
  • Radioactive tracer 33P
  • AMARA peptide

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