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Analysis and interpretation of microplate-based oxygen consumption and pH data.
Methods Enzymol. 547, 309-354 (2014)
Breakthrough technologies to measure cellular oxygen consumption and proton efflux are reigniting the study of cellular energetics by increasing the scope and pace with which discoveries are made. As we learn the variation in metabolism between cell types is large, it is helpful to continually provide additional perspectives and update our roadmap for data interpretation. In that spirit, this chapter provides the following for those conducting microplate-based oxygen consumption experiments: (i) a description of the standard parameters for measuring respiration in intact cells, (ii) a framework for data analysis and normalization, and (iii) examples of measuring respiration in permeabilized cells to follow up results observed with intact cells. Additionally, rate-based measurements of extracellular pH are increasingly used as a qualitative indicator of glycolytic flux. As a resource to help interpret these measurements, this chapter also provides a detailed accounting of proton production during glucose oxidation in the context of plate-based assays.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Bioenergetics ; Ecar ; Glycolysis ; Lactate Efflux ; Mitochondria ; Ocr ; Permeabilized Cells ; Proton Leak ; Respiration ; Seahorse Xf Analyzer
ISSN (print) / ISBN
0076-6879
e-ISSN
0076-6879
Journal
Methods in Enzymology
Quellenangaben
Volume: 547,
Pages: 309-354
Publisher
Elsevier
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of Diabetes and Obesity (IDO)