I have weird declining basal OCR profile while doing a seahorse XFe mitostress experiment. Any suggestions on what is wrong here?

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Ahsan Siraj

Hi Abhinav,

What are these cells? and what about the ECAR levels in this assay? There can be multiple reasons for the drop in OCR (mitochondrial oxidative respiration), for example your cells are changing their phenotype from oxidative to glycolytic respiration in culture which can be seen as drop in OCR. This is true for isolated primary cells like CMs and ECs.

Rafal Czapiewski

It looks like oligomycin might be leaking into the wells or rotenone with AA. You have very small response to oligomycin when it supposed to be observed. Then, really low uncoupled respiration. Check the machine or run it without any chemicals just see if your readout is stable. Hope it helps.