TMRM - Orange Fluorescent Mitochondrial Membrane Depolarization Assay

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TMRM 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 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 2. Jurkat cells were treated with DMSO (control) (A) or staurosporine (B), and then dually stained with TMRM and the green FLICA® poly caspase probe FAM-VAD-FMK (cat. 92) and analyzed by flow cytometer. In the staurosporine-treated cells (B), there is loss of orange fluorescence (FL-2) and a concurrent increase in cell-bound green fluorescence (FL-1) due to caspase activation. Treatment with staurosporine induced apoptosis in about 90% of cells (Ms. Tracy Hanson, ICT).
Figure 1. 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).
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Figure 1. 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 2. Jurkat cells were treated with DMSO (control) (A) or staurosporine (B), and then dually stained with TMRM and the green FLICA® poly caspase probe FAM-VAD-FMK (cat. 92) and analyzed by flow cytometer. In the staurosporine-treated cells (B), there is loss of orange fluorescence (FL-2) and a concurrent increase in cell-bound green fluorescence (FL-1) due to caspase activation. Treatment with staurosporine induced apoptosis in about 90% of cells (Ms. Tracy Hanson, ICT).

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Product Manual
Safety Data Sheet

SKU: 9105

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 TMRM with DMSO.
  5. Dilute MitoPT TMRM with 1X Assay Buffer.
  6. Add MitoPT TMRM 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. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. MitoPT TMRM excites at 548 nm and emits at 573 nm (orange).
  • MitoPT TMRM Reagent, 500 Tests, #6256
  • 10X Assay Buffer, 2 x 125 mL, #6259
  • CCCP, 50 mM, 600 µL, #6258
  • Kit Manual

    • Product Specific References

    PMID Publication
    38871787Wnuk, E, et al. 2024. The physiological levels of epigallocatechin gallate (EGCG) enhance the Cd-induced oxidative stress and apoptosis in CHO-K1 cells. Scientific reports, 13625.
    38768929Shiba-Ishii, A., et al. 2024. Novel therapeutic strategies targeting bypass pathways and mitochondrial dysfunction to combat resistance to RET inhibitors in NSCLC. Biochimica et biophysica acta. Molecular basis of disease, 167249.
    36302969Blanco, D., et al. 2022. PTEN directs developmental and metabolic signaling for innate-like T cell fate and tissue homeostasis. Nature Cell Biology, 1642-1654.
    33627871Lim, S.A., et al. 2021. Lipid signalling enforces functional specialization of Treg cells in tumours. Nature.
    33636132Huang, H., et al. 2021. In vivo CRISPR screening reveals nutrient signaling processes underpinning CD8+ T cell fate decisions. Cell.
    34958665Cai, H., et al. 2021. Critical Role of Lama4 for Hematopoiesis Regeneration and Acute Myeloid Leukemia Progression. Blood.

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