Revision 1
Cell Signaling Technology

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For Research Use Only. Not for Use in Diagnostic Procedures.
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Product Information

Storage

All components in this kit are stable for at least 12 months when stored at the recommended temperature and left unused.
Upon receipt, #9808 should be removed from kit box and stored at room temperature. Remaining components should be stored at -20ºC.

Specificity / Sensitivity

The Mitochondrial Membrane Potential Assay Kit (II) is expected to detect the mitochondrial membrane potential in living cells cross all species. For the best result, a cell number titration is recommended when using a plate-reader and a 96-well plate.

Species Reactivity:

All Species Expected

Product Description

The Mitochondrial Membrane Potential Assay Kit (II) is a fluorescent assay that detects the mitochondrial membrane potential in living cells. The kit includes the cationic dye TMRE (tetramethylrhodamine ethyl ester perchlorate) and a mitochondrial membrane potential disruptor CCCP (carbonyl cyanide 3-chlorophenylhydrazone). TMRE is a cell membrane permeable, fluorescent dye that accumulates in intact mitochondria. Depolarized or inactive mitochondria exhibit decreased membrane potential, resulting in reduced TMRE accumulation.

Background

Mitochondria are the main power house in cells and play important roles in processes such as steroid metabolism, calcium homeostasis, apoptosis and cellular proliferation. Mitochondrial membrane potential is a key indicator of its function and cell health (1,2). The dissipation of mitochondrial membrane potential is established as an early indicator for apoptosis (3).

TMRE (tetramethylrhodamine, ethyl ester) is a cell membrane permeable cationic dye. In normal cells, TMRE accumulates in the mitochondria in response to their high membrane potential and negative charge. When excited at 550 nm, TMRE emits an orange-red fluorescence with a maximum at 575 nm (orange-red). Cells that have lost membrane potential or mitochondria activity cannot accumulate TMRE. Therefore, the fluorescence intensity of the orange-red emission can be used to measure mitochondria membrane potential and is an indicator for cell health (4).

  1. Perry, S.W. et al. (2011) Biotechniques 50, 98-115.
  2. Nesti, C. et al. (2007) Biosci Rep 27, 165-71.
  3. Petit, P.X. et al. (1995) J Cell Biol 130, 157-67.
  4. O'Reilly, C.M. et al. (2003) Biophys J 85, 3350-7.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#13296

Mitochondrial Membrane Potential Assay Kit (II)

Mitochondrial Membrane Potential Assay Kit (II): Image 1 Expand Image
Figure 1. Flow cytometric analysis of Jurkat cells, unlabeled (black) or labeled with 200 nM TMRE and treated with 0 μM CCCP (blue), 3.2 μM CCCP (green), or 80 μM CCCP (red).
Mitochondrial Membrane Potential Assay Kit (II): Image 2 Expand Image
Figure 2. Confocal immunofluorescent analysis of NIH/3T3 cells (2x105 cell/ml) seeded in a 96-well black plate with a clear bottom and incubated overnight. Cells were untreated or treated with CCCP (400 μM, 20 min) followed by labeling with TMRE (200 nM, 30 min).
Mitochondrial Membrane Potential Assay Kit (II): Image 3 Expand Image
Figure 3. HeLa cells (3x105 cell/ml) were treated with various concentrations of CCCP for 15 minutes prior to labeling with 200 nM TMRE.