Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer.

Authors

Colleen L Mayberry, The Jackson LaboratoryFollow
John J Wilson, The Jackson LaboratoryFollow
Britney Sison, The Jackson LaboratoryFollow
Chih-Hao Chang, The Jackson LaboratoryFollow

Document Type

Article

Publication Date

3-25-2024

Original Citation

Mayberry C, Wilson J, Sison B, Chang C. Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer. STAR Protoc. 2024;5(2):102971

Keywords

JMG, SS1

JAX Source

STAR Protoc. 2024;5(2):102971

ISSN

2666-1667

PMID

38536814

DOI

https://doi.org/10.1016/j.xpro.2024.102971

Abstract

Efficient metabolism, or the means by which cells produce energy resources, is critical for proper effector function. Here, we present a protocol for examining the bioenergetics and mitochondrial fuel utilization of primary murine autoreactive immunocytes using cellular metabolism-modulating drugs. We describe steps for plate calibration, isolation of primary immunocytes, and Seahorse assay plate preparation. We then detail procedures for performing the XF Cell Mito Stress Test followed by bioenergetics calculations and statistics. For complete details on the use and execution of this protocol, please refer to Wilson et al.

Comments

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)