|13296||Mitochondrial Membrane Potential Assay Kit (II)||
The CellSimple™ Mitochondrial Membrane Potential Assay Kit (II) is a fluorescent assay designed for use with the CellSimple™ Cell Analyzer. It 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 mean fluorescence intensity (MFI) of the red emission (561 nm LP) can be used as an indicator for mitochondrial membrane potential.
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The CellSimple™ Cell Analyzer is a benchtop instrument that utilizes a disposable thin-film cassette and a combination of a 488 nm laser, two photomultiplier tubes (525/45 nm and 561 nm LP filters), Coulter Principle-based cell measurements, and on-board software to provide easy-to-run applications and data analysis. Data acquisition occurs within approximately 10 seconds per test. The instrument relies on disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and fluidics maintenance and the instrument is small enough to be portable between the lab bench and the hood. Applications include quantitative assessments of cell viability, apoptosis, other labeled antibody markers and single and multiplexed bead-based assays for protein and cellular analysis.
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 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 mitochondrial membrane potential and is an indicator for cell health (4).
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