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  • Isolation and functional analysis of peridroplet mitochondria from murine brown adipose tissue.

Isolation and functional analysis of peridroplet mitochondria from murine brown adipose tissue.

STAR protocols (2021-01-19)
Jennifer Ngo, Ilan Y Benador, Alexandra J Brownstein, Laurent Vergnes, Michaela Veliova, Michael Shum, Rebeca Acín-Pérez, Karen Reue, Orian S Shirihai, Marc Liesa
ABSTRACT

Mitochondria play a central role in lipid metabolism and can bind to lipid droplets. However, the role and functional specialization of the population of peridroplet mitochondria (PDMs) remain unclear, as methods to isolate functional PDMs were not developed until recently. Here, we describe an approach to isolate intact PDMs from murine brown adipose tissue based on their adherence to lipid droplets. PDMs isolated using our approach can be used to study their specialized function by respirometry. For complete information on the use and execution of this protocol, please refer to Benador et al. (2018).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-(−)-Malic acid, BioXtra, ≥95%
Sigma-Aldrich
Potassium phosphate monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%