TMRE - Orange Fluorescent Mitochondrial Membrane Depolarization Assay

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TMRE accumulates and fluoresces orange in polarized mitochondria. Fluorescence drops dramatically on depolarization in apoptotic or metabolically stressed cells. Analyze using a flow cytometer, fluorescence plate reader, or fluorescence microscopy.

Figure 1. Jurkat cells were treated with DMSO (negative control) or 1 µM staurosporine to induce apoptosis, then stained with TMRE, and photographed using a photomicroscope using a green excitation filter at 510-560 nm and a 570-620 nm emission filter (left). Healthy cells, containing mitochondria with polarized inner membranes fluoresce bright orange (A) compared with apoptotic cells with depolarized mitochondria(B). The corresponding DIC images are shown (right) (Dr. Brian Lee, ICT).
Figure 2. Jurkat cells were exposed to DMSO as the negative control (dark orange bars) or 50 µM CCCP depolarizing agent (light orange bars) and then incubated with TMRE or TMRM. Aliquots (100 µL) were analyzed in triplicate in a black 96-well plate using a fluorescence plate reader (550 nm excitation and 580 nm emission using a 570 nm cut-off filter). Healthy control cells had a high level of orange fluorescence compared to the CCCP-treated cells. (Ms. Tracy Hanson, ICT).
Figure 3. Jurkat cells were treated with 1 µM staurosporine, an apoptosis-inducing agent (left, white), or DMSO, a negative control (right, orange), then stained with TMRE for 15-20 minutes Cells were then analyzed by flow cytometer. Apoptotic cells (left), with depolarized mitochondria, exhibit much less orange fluorescence compared to healthy cells with polarized mitochondrial inner membranes; these cells concentrate TMRE and fluoresce bright orange. (Ms. Tracy Hanson, ICT).
Figure 1. Jurkat cells were treated with DMSO (negative control) or 1 µM staurosporine to induce apoptosis, then stained with TMRE, and photographed using a photomicroscope using a green excitation filter at 510-560 nm and a 570-620 nm emission filter (left). Healthy cells, containing mitochondria with polarized inner membranes fluoresce bright orange (A) compared with apoptotic cells with depolarized mitochondria(B). The corresponding DIC images are shown (right) (Dr. Brian Lee, ICT).
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Figure 1. Jurkat cells were treated with DMSO (negative control) or 1 µM staurosporine to induce apoptosis, then stained with TMRE, and photographed using a photomicroscope using a green excitation filter at 510-560 nm and a 570-620 nm emission filter (left). Healthy cells, containing mitochondria with polarized inner membranes fluoresce bright orange (A) compared with apoptotic cells with depolarized mitochondria(B). The corresponding DIC images are shown (right) (Dr. Brian Lee, ICT).
Figure 2. Jurkat cells were exposed to DMSO as the negative control (dark orange bars) or 50 µM CCCP depolarizing agent (light orange bars) and then incubated with TMRE or TMRM. Aliquots (100 µL) were analyzed in triplicate in a black 96-well plate using a fluorescence plate reader (550 nm excitation and 580 nm emission using a 570 nm cut-off filter). Healthy control cells had a high level of orange fluorescence compared to the CCCP-treated cells. (Ms. Tracy Hanson, ICT).
Figure 3. Jurkat cells were treated with 1 µM staurosporine, an apoptosis-inducing agent (left, white), or DMSO, a negative control (right, orange), then stained with TMRE for 15-20 minutes Cells were then analyzed by flow cytometer. Apoptotic cells (left), with depolarized mitochondria, exhibit much less orange fluorescence compared to healthy cells with polarized mitochondrial inner membranes; these cells concentrate TMRE and fluoresce bright orange. (Ms. Tracy Hanson, ICT).

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Product Manual
Safety Data Sheet

SKU: 9103

Size: 500 Tests
Price:
$299.00
Ships: 1-2 business days
  1. Prepare samples.
  2. Create controls with CCCP.
  3. Dilute 10X Assay Buffer 1:10 with diH2O.
  4. Reconstitute MitoPT TMRE with DMSO.
  5. Dilute MitoPT TMRE with 1X Assay Buffer.
  6. Add MitoPT TMRE to each sample.
  7. Incubate 15-30 minutes.
  8. Wash cells: add 1X Assay Buffer and spin cells.
  9. Remove supernatant and resuspend cells for analysis.
  10. Anazlyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. MitoPT TMRE excites at 549 nm and emits at 574 nm (orange).
  • MitoPT TMRE Reagent, 500 Tests, #6254
  • 10X Assay Buffer, 2 x 125 mL, #6259
  • CCCP, 50 mM, 600 µL, #6258
  • Kit Manual

    • Product Specific References

    PMID Publication
    39125651Nukaga, S., et al. 2024. Caprylic Acid Inhibits High Mobility Group Box-1-Induced Mitochondrial Damage in Myocardial Tubes. International Journal of Molecular Sciences, .
    39123382Montagnani Marelli, M, et al. 2024. Anticancer Activity of Delta-Tocotrienol in Human Hepatocarcinoma: Involvement of Autophagy Induction. Cancers, 2654.
    38247469Günther, M., et al. 2023. Flavonoid-Phenolic Acid Hybrids Are Potent Inhibitors of Ferroptosis via Attenuation of Mitochondrial Impairment. Antioxidants, 44.
    37627584Merkel, M., et al. 2023. Mitochondrial Reactive Oxygen Species Formation Determines ACSL4/LPCAT2-Mediated Ferroptosis. Antioxidants (Basel, Switzerland).
    37760171Asadipour, K., et al. 2023. Ultra-Low Intensity Post-Pulse Affects Cellular Responses Caused by Nanosecond Pulsed Electric Fields. Bioengineering (Basel, Switzerland).
    28981113Rehklau, K., et al. 2017. Cofilin1-dependent actin dynamics control DRP1-mediated mitochondrial fission. Cell Death Discovery, e3063.

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