Buy a SimpleChIP kit and receive a sample of our new Universal qPCR Master Mix* | Learn More >>
13296
Mitochondrial Membrane Potential Assay Kit (II)

Mitochondrial Membrane Potential Assay Kit (II) #13296

Flow Cytometry Image 1

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).

Learn more about how we get our images
Fluorescent Detection Image 2

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).

Learn more about how we get our images
Cell-based assay Image 3

Figure 3. HeLa cells (3x105 cell/ml) were treated with various concentrations of CCCP for 15 minutes prior to labeling with 200 nM TMRE.

Learn more about how we get our images
Product Includes Quantity (with Count) Solution Color
TMRE 1 x 29 µg Purple
CCCP 1 x 100 µl Yellow
Phosphate Buffered Saline (PBS-20X) 9808 1 x 25 ml Colorless

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.

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.

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.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

News from the Bench

Discover what’s going on at CST, receive our latest application notes and tips, read our science features, and learn about our products.

Subscribe

To Purchase # 13296S

Product Number Size Price
13296S 1 Kit (500 assays (96 well format)) $265.00.0
Quantity Subtotal
$0.00