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

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

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10.1016/j.xpro.2021.100730

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title = "Isolation of autophagic fractions from mouse liver for biochemical analyses",

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).",

keywords = "cell biology, cell separation/fractionation, metabolism",

author = "Henrietta Bains and Rajat Singh",

note = "Funding Information: This work was supported by 6R01AG043517-05 (R.S.), R01DK123327 (R.S.), 5P01AG031782 (Project leader, R.S.), R01AG065985-01 (R.S.), and ADA Grant 1-18-IBS-062 (R.S.). H.B. is supported by NIA 1F31AG071112-01 . We thank Dr. Susmita Kaushik (Research Assistant Professor, Developmental and Molecular Biology, Albert Einstein College of Medicine) for her helpful suggestions. Cartoon depicting scheme for isolation of autophagic fractions were made with Biorender.com . Publisher Copyright: {\textcopyright} 2021 The Author(s)",

year = "2021",

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doi = "10.1016/j.xpro.2021.100730",

language = "English (US)",

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T1 - Isolation of autophagic fractions from mouse liver for biochemical analyses

AU - Bains, Henrietta

AU - Singh, Rajat

N1 - Funding Information: This work was supported by 6R01AG043517-05 (R.S.), R01DK123327 (R.S.), 5P01AG031782 (Project leader, R.S.), R01AG065985-01 (R.S.), and ADA Grant 1-18-IBS-062 (R.S.). H.B. is supported by NIA 1F31AG071112-01 . We thank Dr. Susmita Kaushik (Research Assistant Professor, Developmental and Molecular Biology, Albert Einstein College of Medicine) for her helpful suggestions. Cartoon depicting scheme for isolation of autophagic fractions were made with Biorender.com . Publisher Copyright: © 2021 The Author(s)

PY - 2021/9/17

Y1 - 2021/9/17

N2 - 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).

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).

KW - cell biology

KW - cell separation/fractionation

KW - metabolism

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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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