Isolation of autophagic fractions from mouse liver for biochemical analyses

Henrietta Bains; Rajat Singh

doi:10.1016/j.xpro.2021.100730

Isolation of autophagic fractions from mouse liver for biochemical analyses

Henrietta Bains, Rajat Singh

Medicine

Research output: Contribution to journal › Article › peer-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 language	English (US)
Article number	100730
Journal	STAR Protocols
Volume	2
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2021.100730
State	Published - Sep 17 2021

Keywords

cell biology
cell separation/fractionation
metabolism

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Neuroscience
General Immunology and Microbiology
General Medicine

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10.1016/j.xpro.2021.100730

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AU - Singh, Rajat

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AB - 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).

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KW - cell separation/fractionation

KW - metabolism

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JF - STAR Protocols

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ER -

Isolation of autophagic fractions from mouse liver for biochemical analyses

Abstract

Keywords

ASJC Scopus subject areas

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