Isolation and analysis of fractions enriched in WrappER-associated mitochondria from mouse liver

Abstract

Summary: The endoplasmic reticulum (ER) plays a central role in lipid homeostasis, but the role of individual ER subdomains in lipid biology has not been elucidated. WrappER is a curved wrapping type of rough-ER that establishes extensive contacts with almost every mitochondria of the hepatocyte in the mouse liver. Here, we describe a protocol for isolation of fractions enriched in wrappER-associated mitochondria from the mouse liver. We also provide techniques for assessing its quality by electron microscopy and biochemical/proteomic analysis.For complete information on the use and execution of this protocol, please refer to Anastasia et al. (2021).

Keywords

• Cell Biology
• Cell separation/fractionation
• Metabolism
• Protein Biochemistry