Hello, I performed a titration of FCCP in HaCaT cells in order to do a mito stress test with the Seahorse technology (Agilant make). I have tried 0.125, 0.25µM, 0.5µM, 1 and 2µM concentration of FCCP but can't see an increase in OCR (even bellow basal level). Has anyone encountered this situation with HaCaT cells? and what can be done about it?
Thank you!
Hi Hemang,
I see the same phenomenon when I take measurements of HeLa cells in the Seahorse. Have you been able to solve this yet?
I could slightly increase the the OCR as a reaction to FCCP in Hela cells when I add 1% FCS into the assay (at least to the point where the max respiration is slightly higher than the basal).
Thanks,
Juliane
Hi @juliane,
Yes.... Finally, I optimized the experiment.
I would suggest to start with lower concentration of FCCP (try with 0.01uM). Some cells are too sensitive to FCCP. At higher concentrations, they got saturated.
Best of luck for the experiment.
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