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Seahorse analysis

AM Ashley A. Viehmann Milam
JB Juliet M. Bartleson
MB Michael D. Buck
CC Chih-Hao Chang
AS Alexey Sergushichev
DD David L. Donermeyer
WL Wing Y. Lam
EP Erika L. Pearce
MA Maxim N. Artyomov
PA Paul M. Allen
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OCR and ECAR were measured using a 96-well XF extracelluar flux analyzer (Seahorse Bioscience) and Extracellular Flux Assay Kit (Agilent Seahorse XFe96). Assay setup has been previously described (30–33). Briefly, cells were plated in XF media supplemented with glucose (25mM), L-glutamine (2mM), and sodium pyruvate (1mM). Measurements were taken at basal state, and after the step-wise addition of oligomycin (1μM), FCCP (1.5μM), and rotenone (100μM) plus antimycin A (1μM). The mGPD2 specific inhibitor, iGP-1 (Calbiochem/EMD Millipore #5.30655.0001) was dissolved in DMSO and added to the extracellular flux analyses at either 2.5μM or 5.0μM final concentration.

Copyright and License information:  ©2020-08-07

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