Isolation of autophagic fractions from mouse liver for biochemical analyses

Henrietta Bains, Rajat Singh

Research output: Contribution to journalArticlepeer-review

Abstract

Isolation of autophagosomes, autolysosomes, and lysosomes allows mechanistic studies into the pathophysiology of autophagy—a lysosomal quality control pathway. Here, we outline a Nycodenz density gradient ultracentrifugation approach for high-yield isolation of autophagic fractions from mouse liver. These fractions can be used for immunoblotting, transmission electron microscopy, and proteomic and lipidomic analyses. For complete details on the use and execution of this protocol, please refer to Toledo et al. (2018).

Original languageEnglish (US)
Article number100730
JournalSTAR Protocols
Volume2
Issue number3
DOIs
StatePublished - Sep 17 2021

Keywords

  • cell biology
  • cell separation/fractionation
  • metabolism

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)

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