Abstract
Interest in the investigation of mitochondrial dysfunction has seen a resurgence over recent years due to the implication of such dysfunction in both drug-induced toxicity and a variety of disease states. Here we describe a methodology to assist in such investigations whereby the oxygen consumption of isolated mitochondria is assessed in a high-throughput fashion using a phosphorescent oxygen-sensitive probe , standard microtiter plates, and plate reader detection. The protocols provided describe the required isolation procedures, initial assay optimization, and subsequent compound screening. Typical data is also provided illustrating the expected activity levels as well as recommended plate maps and data analysis approaches.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Dykens JA, Marroquin LD, Will Y (2007) Strategies to reduce late-stage drug attrition due to mitochondrial toxicity. Expert Rev Mol Diagn 7:161–175
Wallace KB (2008) Mitochondrial off targets of drug therapy. Trends Pharmacol Sci 29:361–366
Papkovsky DB, Hynes J, Will Y (2006) Respirometric screening technology for ADME-Tox studies. Expert Opin Drug Metab Toxicol 2:313–323
Will Y, Hynes J, Ogurtsov VI, Papkovsky DB (2007) Analysis of mitochondrial function using phosphorescent oxygen-sensitive probes. Nat Protoc 1:2563–2572
Hynes J, Hill R, Papkovsky DB (2006) The use of a fluorescence-based oxygen uptake assay in the analysis of cytotoxicity. Toxicol In Vitro 20:785–792
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
1 Electronic Supplementary Material
Supplemental Data 1:
Data Transformation and calculation of UC50 and IC50 data (DOC 3676 kb)
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Hynes, J., Swiss, R.L., Will, Y. (2018). High-Throughput Analysis of Mitochondrial Oxygen Consumption. In: Palmeira, C., Moreno, A. (eds) Mitochondrial Bioenergetics. Methods in Molecular Biology, vol 1782. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7831-1_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7831-1_4
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7830-4
Online ISBN: 978-1-4939-7831-1
eBook Packages: Springer Protocols