• Product name
    TMRE-Mitochondrial Membrane Potential Assay Kit
    See all Mitochondrial Membrane Potential kits
  • Detection method
  • Sample type
    Adherent cells, Suspension cells
  • Assay type
    Cell-based (qualitative)
  • Assay time
    0h 35m
  • Product overview

    TMRE-Mitochondrial Membrane Potential Assay Kit ab113852 is used for quantifying changes in mitochondrial membrane potential in live cells by flow cytometry, microplate spectrophotometry and fluorescent microscopy. 

    ab113852 uses TMRE (tetramethylrhodamine, ethyl ester) to label active mitochondria. TMRE is a cell permeant, positively-charged, red-orange dye that readily accumulates in active mitochondria due to their relative negative charge. Depolarized or inactive mitochondria have decreased membrane potential and fail to sequester TMRE.

    The TMRE protocol also uses FCCP (carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone), which is a ionophore uncoupler of oxidative phosphorylation. Treating cells with FCCP eliminates mitochondrial membrane potential and TMRE staining. TMRE is suitable for the labeling of mitochondria in live cells and is not compatible with fixation.

    TMRE protocol summary:
    - add FCCP to appropriate control cell samples and incubate for 10 min
    - incubate with TMRE for 15-30 min, pellet (suspension cells) / remove media (adherent cells) and wash with PBS / 0.2% BSA
    - analyze with micro-plate reader at Ex/Em 549/575 nm, flow cytometer using 488nm laser for excitation and at emission 575 nm, or fluorescent microscope.

  • Notes

    TMRE is only suitable for use with live (not fixed) cells.

    Related assays

    Review the cell health assay guide to learn about kits to perform a cell viability assay, cytotoxicity assay and cell proliferation assay

    Review the metabolism assay guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

  • Platform
    Microplate reader, Fluor. microscope, Flow cyt.



  • P19 neurons (750 cells/mm2) were exposed to MDMA on days 7–9 in serum-free medium for 10 min up to 48 hours. The positive control FCCP (carbonyl cyanide-p-trifluoromethoxyphenylhydrazone), an uncoupler of mitochondrial oxidative phosphorylation, was applied at the concentration of 5 μM for 10 min. The cells were incubated with 500 μM TMRE for 30–45 min at 37°C, 5% CO2, followed by washing once with 100 μl of HBSS containing 0.2% bovine serum albumin. A volume of 200 μL of HBSS containing 0.2% bovine serum albumin was added to each well, and the fluorescence was measured with excitation/emission: 544/590 nm.

  • A: HeLa cells (adherent) were cultured on coverslips and stained with ab113852 (200nM TMRE) for 20 minutes in media, washed briefly with PBS and immediately imaged. B: Jurkat cells (suspension) were stained and washed as above and then transferred to a slide and immobilized under a coverslip for imaging.
  • Chart showing mean fluorescent intensity +/- standard deviation from quadruplicate measurements of 400 nM TMRE stained Jurkat cells in a 96-well microplate +/- treatment with FCCP.

  • Flow cytometry histogram of Jurkat cells stained with ab113852 (100nM TMRE) with (blue) or without (red) treatment with 100µM FCCP.



This product has been referenced in:
  • Lee HA  et al. Histone deacetylase inhibitor MGCD0103 protects the pancreas from streptozotocin-induced oxidative stress and ß-cell death. Biomed Pharmacother 109:921-929 (2019). Read more (PubMed: 30551546) »
  • Yoon YM  et al. TUDCA-treated chronic kidney disease-derived hMSCs improve therapeutic efficacy in ischemic disease via PrPC. Redox Biol 22:101144 (2019). Read more (PubMed: 30785084) »
See all 60 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A


Thank you for contacting us. Our lab uses:

BD PureCoat™ Microplates
black/clear, polystyrene
96-well, amine

The recommendation is for clear bottom, black wall microplate that (a) cells will grow on and (b) is compatible with plate reader.

Please let me know if you have any other questions.

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Thank you for your inquiry. I heard back from the lab regardingthese two distinct dyes that both measure mitochondrial membrane potential. TMRE stains mitochondria only when there is a membrane potential. JC-1 is slightly different in that it has distinct emission spectra depending on whether mitochondria have high or low potential. TMRE can be measured in a spectophotometer, fluorescent microscope or flow cytometry. JC-1 can be measured in a spectrophotometer. Details are in the respective protocol booklets. https://www.abcam.com/ps/products/113/ab113850/documents/ab113850%20Protocol%20(Website)%20v2.pdf https://www.abcam.com/ps/products/113/ab113852/documents/ab113852%20protocol%20final%20v2%20(website(.pdf For you, wewould recommend TMRE with analysis on a flow cytometer. Flow cytometry is both sensitive and amenable to very few cells. I hope this information helps. Please contact us with any other questions.

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To measure mitochondrial activity in human fibroblast cells, we applied TMRE mitochondrial membrane potential assay kit. It showed great correlation with Seahorse analysis which measured OCR (oxygen consumption rate).

Abcam user community

Verified customer

Submitted May 02 2018


The samples should be read immediately after the 15-30 minute incubation with TMRE. Since the cells are live, as the health of the cell declined the signal from the dye would also diminish.

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For another positive control, CCCP is suitable (5-50 μM CCCP for 30 to 60 minutes at 37ºC).

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It all depends on the sample (embryos vs. explant vs. whole work). We have only worked with this dye using tissue culture cells (monolayer or suspension). If the samples are dissociated cells then TMRE labeling should work. Otherwise, we don’t know how well or deeply the dye can penetrate across cell layers (e.g. into tissues or embryos).

The dye for measuring the potential in the kit is species independent, so it is more a matter of the sample type.

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We have no experience in using TMRE to stain isolated mitochondria from any source – it is a tricky proposition as you would need intact/non-damaged mitochondria which retain membrane potential therefore we wouldn't recommend this.

As for imaging tissue stained with TMRE, this is also tricky. Flow cytometry is almost certainly out due to the need for single cell suspension – which will be very difficult for most tissue types. For microscopy I suppose it is possible with the proper microscope set up and the ability to adapt the live tissue to microscope stage constraints (e.g. thin slices.)

However, I would like to stress that this kit is only really for mitochondria labelling in intact cultured cells.

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Thank you for contacting us. My colleague from our MitoSciences lab has provided the following recommendations.

The lack of signal may be due to working outside the peak excitation (549nm) and peak emission (575nm). Getting as close as possible to these parameters is critical for maximal signal.

Many fluorescent spectrophotometers read only in the center of the well. It may help to ensure that your plate reader is capable of reading at the bottom of the plate and ensure the instrument is set up as such. We would also recommend to increase the PMT gain (photomultiplier tube) on the instrument.

Another option is to increase the concentration of TMRE to 1000nM and increase the density of the cells. With mouse embryonic fibroblasts, we would recommend to test the following densities on a 96 well plate to find optimal seeding = 50,000 / 25,000 / 10,000 / 5,000. Using a titration of cells, we recommend testing different TMRE concentrations (1000nM, 200nM, 50nM).

I hope this helps, if not, please let me know and I will be happy to help you further.

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Thank you for contacting us.

I have received the following information from the lab:

TMRE can only be used with live cells—an active membrane potential is required for staining. This will be true of all membrane potential sensitive dyes (including JC-1).

As for assessing mitochondria in the existing samples:
One possibility is to look at Cytochrome C release from the mitochondria by ICC, e.g. the mouse monoclonal ab110325. (see also Figure 11 in the MitoSciences Apoptosis Playbook here: http://mitosciences.com/PDF/Apoptosis_Playbook.pdf

Another option might be the ApoTrack™ Cytochrome c Apoptosis ICC Antibody Kit (ab110417)

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Thank you for your inquiry.

Culture media is known to cause background using kit ab113852. For suspension cells, we recommend pelleting the cells, removing the culture media, resuspending in the same volume of 0.2% BSA in PBS, pelleting again, and resuspending a second time in 0.2% BSA in PBS before transferring to a microplate.

I hope this information helps. Please contact us with any other questions.

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1-10 of 17 Abreviews or Q&A

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