Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
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

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

All Species Expected

Product Description

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.

Background

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).

  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
#45898

CellSimple Mitochondrial Membrane Potential Assay Kit (II)

CellSimple™ Mitochondrial Membrane Potential Assay Kit (II): Image 1 Expand Image
CellSimple cell-based analysis of live Jurkat cells that were labeled with TMRE (200 nM, 30 min) and untreated (left panel) or CCCP-treated (50 μM, 37ºC, 15 min; right panel) using the CellSimple Mitochondrial Membrane Potential Assay Kit (II) #45898. Data was collected in both red (561 nm LP) and green (525/45 nm) channels and analyzed on the Open Flow Cytometry application. Instrument screen shots are shown.
CellSimple™ Mitochondrial Membrane Potential Assay Kit (II): Image 2 Expand Image
Mean Fluorescence Intensity (MFI) of TMRE labeled (200 nM, 30 min) live Jurkat cells untreated (black) or treated with CCCP (50 μM, 37ºC, 15 min; blue). The 'overlay file' feature on the CellSimple instrument is used to display MFI in a histogram format. The shift in MFI in the red (561nm LP) channel demonstrates the change in membrane potential.