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Assessing Calcium-Stimulated Mitochondrial Bioenergetics Using the Seahorse XF96 Analyzer

  • Jennifer Wettmarshausen
  • Fabiana PerocchiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1925)

Abstract

The development of fluorescence-based oxygen sensors coupled with microplate-based assays for quantitative bioenergetics analyses enables screening multiple experimental conditions at once with small biological material and in a timely manner. In this chapter, we outline detailed protocols and practical tips to design and perform controlled measurements of (a) respiratory and glycolytic metabolism of intact cells, (b) substrate-dependent respiration in permeabilized cells and isolated mitochondria, and (c) calcium-dependent regulation of mitochondrial bioenergetics with Seahorse XF Flux Analyzers.

Key words

Mitochondria Bioenergetics Respiration Oxygen consumption Intact cells Permeabilized cells Seahorse XF analyzer Calcium 

Notes

Acknowledgments

We thank Dr. Daniela Arduino and Valerie Goh for critical reading of the manuscript. This work was supported by the German Research Foundation (DFG) under the Emmy Noether Programme (PE 2053/1-1) and the Bert L & N Kuggie Vallee Foundation (to F.P. and J.W.).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute for Diabetes and Obesity, Helmholtz Diabetes Center (HDC), Helmholtz Zentrum München and German National Diabetes Center (DZD)NeuherbergGermany
  2. 2.Department of BiochemistryGene Center Munich, Ludwig-Maximilians Universität MünchenMunichGermany

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