Identifying the heterotrimeric complex stoichiometry of AMPK in skeletal muscle by immunoprecipitation

1 Citation (Scopus)

Abstract

The 5'-AMP-activated protein kinase is a complicated enzyme consisting of three different subunits, each of which is expressed as two or three isoforms. This gives the possibility of 12 different heterotrimeric complexes, which could have diverse functions within the cell. To map out which of these complexes are present and to what extent in skeletal muscle, we have used the immunoprecipitation technique and analyzed both the precipitates and the remaining supernatants for coprecipitation of complex partners. We have fine-tuned this method to give the best results on lysates from the skeletal muscle, liver, and heart muscle from mouse to man.

Original languageEnglish
Title of host publicationAMPK : Methods and Protecols
EditorsDietbert Neumann, Benoit Viollet
Number of pages11
Place of PublicationNew York
PublisherHumana Press
Publication date2018
Pages203-213
Chapter13
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
  • AMPK
  • Homogenization
  • Protein interaction
  • Immunoprecipitation
  • Western blot
  • In vitro setting
  • Antibody specificity

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