MitoProbe™ TMRM Assay Kit for Flow Cytometry - FAQs

Yes. The CCCP should be first added to the cell sample, incubated for 5 mins, and then the TMRM should be added and incubated for another 30 mins.

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The Image-iT TMRM Reagent is provided as a 100 µM solution of TMRM in DMSO (5 vials, 100 µL in each vial). The MitoProbe TMRM Assay Kit for Flow Cytometry provides the TMRM as a 20 µM stock solution in DMSO (1 vial, 100 µL) and a vial of CCCP. Both kits have the stock solutions as a 1000X concentration, but final working concentrations required for imaging applications are 5-fold higher than what is needed for flow cytometry analysis.

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TMRM is a dynamic mitochondrial stain. There is no covalent attachment of TMRM to the mitochondria and because of this, it can move in and out of the mitochondria dependent upon changes in the membrane potential. The MitoTracker dyes on the other hand possess a reactive chloromethyl group that allows for their covalent attachment to mitochondria. As a result, once they accumulate within mitochondria that have a normal membrane potential, they are retained.

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