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

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 stab...

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Detalles Bibliográficos
Autores principales: Bose, Himangshu S., Doetch, Nicole E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9841272/
https://www.ncbi.nlm.nih.gov/pubmed/36620990
http://dx.doi.org/10.1016/j.xpro.2022.101996
Descripción
Sumario: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)

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