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

CellSimple Mitochondrial Membrane Potential Assay Kit (I) 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 JC-1 and a mitochondrial membrane potential disruptor CCCP (carbonyl cyanide 3-chlorophenylhydrazone). JC-1 is a cell membrane permeable, fluorescent dye with green emission (~520 nm). When JC-1 accumulates in intact mitochondria, the dye forms aggregates that lead to orange-red fluorescence (~590 nm). The mean fluorescence intensity (MFI) of the orange-red emission 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 function as the main cellular powerhouse and play important roles in other processes, such as steroid metabolism, calcium homeostasis, apoptosis, and cellular proliferation. Mitochondrial membrane potential is a key indicator of mitochondrial function and cell health (1,2). The dissipation of mitochondrial membrane potential is considered an early indicator of apoptosis (3).

JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide) is a cell membrane permeable, cationic dye. In normal cells, JC-1 concentrates in mitochondria to form aggregates in response to high membrane potential. Decreased mitochondrial membrane potential results in dispersal of mostly monomeric JC-1 throughout the cell. When excited at 490 nm, JC-1 monomers emit a green fluorescence with a maximum at ~520 nm. Aggregates of JC-1 emit an orange-red fluorescence with a maximum at ~590 nm. Therefore, the fluorescence intensity of the orange-red emission and the ratio of orange-red fluorescence to green fluorescence can be used to measure mitochondrial membrane potential and serve as an indicator of overall 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. Perelman, A. et al. (2012) Cell Death Dis 3, e430.

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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 1
#42239

CellSimple Mitochondrial Membrane Potential Assay Kit (I)

CellSimple™ Mitochondrial Membrane Potential Assay Kit (I): Image 1 Expand Image
CellSimple cell-based analysis of live Jurkat cells untreated (left panel) or CCCP-treated (50 μM, 37ºC, 15 min; right panel) and labeled with JC-1 (2 μM, 30 min) using the CellSimple Mitochondrial Membrane Potential Assay Kit (I). 采集红色 (561 nm LP) 和绿色 (525/45 nm) 通道的数据,并在 Open Flow Cytometry 应用上进行分析。在 CCCP 处理时,注意红色通道中明显下降的平均荧光强度。仪器屏幕截图如图所示。