Investigating the K⁺ sensitivity of cellular metabolism by extracellular flux analysis

We have recently demonstrated that the activity of hexokinase 2 is dependent on the intracellular potassium ion (K⁺) concentration ([K⁺]). To analyze the K⁺ dependency of the cell metabolism in cell populations, we used an extracellular flux analyzer to assess oxygen consumption and acidification rates as well-established measures of oxidative- and glycolytic metabolic activities. This protocol describes in detail how a potential K⁺ sensitivity of the cell metabolism can be elucidated by extracellular flux analysis. For complete details on the use and execution of this protocol, please refer to Bischof et al. (2021).