Protocol for direct measurement of stability and activity of mitochondria electron transport chain complex II

Abstract

Summary: Mitochondria electron transport chain (ETC) complex II is essential for steroid metabolism. Here, we present a protocol to measure the stability and activity of mitochondria ETC complex II. We first describe mitochondria isolation from cell lines and tissues. We then detail how to determine the stability of ETC complex II using isothermal calorimetry and quantification of steroidogenesis using activity assays in parallel. Finally, we describe the steps to perform radioimmunoassay (RIA) to confirm the activity of ETC complex II.For complete details on the use and execution of this protocol, please refer to Bose et al. (2020).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Isolation
• Cell-based Assays
• Metabolism
• Protein Biochemistry