Journal Article Research Support, Non-U.S. Gov't Research Support, N.I.H., Extramural Ppublish 1 citation
PubMed: 38850537 DOI: 10.1016/j.xpro.2024.103042
Protocol for Seahorse analysis of ex vivo mouse brown and white adipose tissues.
STAR Protoc 5(2):103042
Authors: Fenfen Wang, Phu M Huynh, Yu A An
Affiliations
- Department of Anesthesiology, Critical Care and Pain Medicine, Center for Perioperative Medicine, McGovern Medical School, UT Health Science Center at Houston, Houston, TX 77030, USA.
- Department of Anesthesiology, Critical Care and Pain Medicine, Center for Perioperative Medicine, McGovern Medical School, UT Health Science Center at Houston, Houston, TX 77030, USA.
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-CO 2 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.
MeSH
Stress, Physiological, Clinical Laboratory Techniques, Tissues
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