Measurement of mitochondrial respiration in adherent cells by Seahorse XF96 Cell Mito Stress Test

Mitochondria play pivotal roles in cellular energy metabolism. Most of the intracellular adenosine triphosphate (ATP) is generated by mitochondrial respiration. The Cell Mito Stress Test is a common method to measure the key parameters of mitochondrial respiration. Here, we use the human cell line HK-2 as an example to present the procedures to quantify the oxygen consumption rate using a Seahorse XFe96 extracellular flux analyzer. For complete details on the use and execution of this protocol, please refer to Ma et al. (2020).