Protocol for Seahorse Analysis of Ex Vivo Mouse Brown and White Adipose Tissues

Authors

Fenfen Wang
Phu M Huynh
Yu A An

Publication Date

6-21-2024

Journal

Star Protocols

Abstract

The mitochondrial stress test is a gold-standard approach for assessing adipose tissue physiological functions and pathological changes. Here, we present a protocol for conducting Seahorse assays using ex vivo mouse brown and white adipose depots. We describe steps for rehydrating the cartridge, preparing freshly harvested fat depots, placing them onto an islet capture plate, and incubating them in a non-CO2 incubator. We then detail procedures for adding mitochondrial stressor solutions and conducting the mitochondrial stress test using the Seahorse XFe24 Analyzer. For complete details on the use and execution of this protocol, please refer to An et al.1.

Keywords

Animals, Mice, Adipose Tissue, White, Adipose Tissue, Brown, Mitochondria, Cell-based Assays, Health Sciences, Metabolism

Comments

PMID: 38850537