The MitoPT TMRM Assay detects mitochondrial membrane depolarization utilizing the fluorescent dye TMRM. When accumulated in a negatively charge polarized mitochondria, TMRM fluoresces orange. When mitochondrial membrane potential collapses in apoptotic or metabolically stressed cells, TMRM reagent is dispersed through the cell cytosol and fluorescence levels drop dramatically. Analyze your results using a flow cytometer, fluorescence plate reader, or fluorescence microscopy.

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MitoPT TMRM Assay
SKU: 9105

Size: 500 Tests
Price:
Sale price$270


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Loss of mitochondrial membrane potential is indicative of apoptosis. ICT’s MitoPT TMRM Assay utilizes the potentiometric fluorescent dye TMRM to detect mitochondrial membrane permeability and membrane depolarization. The TMRM dye has a delocalized positive charge dispersed throughout its molecular structure. In addition, its lipophilic solubility enables it to be readily membrane permeant and penetrate living cells. The TMRM dye enters the negatively charge mitochondria where it accumulates and fluoresces orange upon excitation. When the mitochondrial membrane potential collapses in apoptotic cells, MitoPT TMRM becomes distributed throughout the cytosol. Detection of the loss of orange-red fluorescence in TMRM stained cells is a reliable method for assessing apoptosis induction or oxidative stress-induced mitochondrial depolarization in experimental cell populations.
TMRM
Mitochondrial depolarization
548 nm / 573 nm
Flow cytometry, Fluorescence microscope, fluorescence plate reader
Cell culture
-20°C
Domestic: Overnight Delivery; International: Priority Shipping
United States
  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
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    Wei J, Long L, Zheng W, Dhungana Y, Lim SA, Guy C, Wang Y, Wang YD, Qian C, Xu B, Kc A, Saravia J, Huang H, Yu J, Doench JG, Geiger TL, Chi H. Targeting REGNASE-1 programs long-lived effector T cells for cancer therapy. Nature. 2019 Dec;576(7787):471-476. doi: 10.1038/s41586-019-1821-z. Epub 2019 Dec 11. Abstract

    Raynor JL, Liu C, Dhungana Y, Guy C, Chapman NM, Shi H, Neale G, Sesaki H, Chi H. Hippo/Mst signaling coordinates cellular quiescence with terminal maturation in iNKT cell development and fate decisions. J Exp Med. 2020 Jun 1;217(6):e20191157. doi: 10.1084/jem.20191157. Abstract

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