Using Seahorse Machine to Measure OCR and ECAR in Cancer Cells

  • Jing ZhangEmail author
  • Qing ZhangEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1928)


A large amount of energy used for nutrient processing and cellular functions is essential for tumorigenesis. Total intracellular adenosine triphosphate (ATP) is mainly generated by glycolysis and mitochondrial oxidative phosphorylation. Here, we provide a protocol for measurements of energy metabolism in cancer cells by using Seahorse XF24 Extracellular Flux analyzer. Specifically, this machine measures glycolysis by analyzing the extracellular acidification rate (ECAR) and measures mitochondrial oxidative phosphorylation on the basis of the oxygen consumption rate (OCR), through real-time and live cell analysis. This protocol is provided for researchers who are unfamiliar with the method and to aid them in carrying out the technique successfully.

Key words

Energy metabolism Glycolysis Oxidative phosphorylation Seahorse XF24 Extracellular Flux analyzer OCR ECAR 



We thank Yong Liu and Agilent agents for helpful discussions and suggestions. Jing Zhang is supported by a DoD BCRP Breakthrough Fellowship Award (W81XWH-17-1-0016). Qing Zhang is supported by grants from the National Cancer Institute (R01CA211732, R21CA223675) and American Cancer Society (RSG-18-059-01-TBE). Qing Zhang also received a career development award from the DoD (W81XWH-15-1-0599). Qing Zhang is a V Scholar, Kimmel Scholar, Komen Career Catalyst Awardee, and Mary Kay Foundation Awardee.


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Copyright information

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

Authors and Affiliations

  1. 1.Department of Pathology and Laboratory MedicineLineberger Comprehensive Cancer Center, UNC-Chapel HillChapel HillUSA
  2. 2.Department of PharmacologyUNC-Chapel HillChapel HillUSA

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