logo
AAT Bioquest

Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry Assays*

Effect of FCCP induced mitochondria membrane potential change in Jurkat cells. Jurkat cells were dye loaded with JC-10 dye working solution along with DMSO (Top) or 5 µM FCCP (Low) for 10 minutes. The fluorescence intensities for both J-aggregates and monomeric forms of JC-10 were measured with a FACSCalibur (Becton Dickinson) flow cytometer using FL1 and FL2 channels. Uncompensated data (left column) were compared with compensated data (right column).
Effect of FCCP induced mitochondria membrane potential change in Jurkat cells. Jurkat cells were dye loaded with JC-10 dye working solution along with DMSO (Top) or 5 µM FCCP (Low) for 10 minutes. The fluorescence intensities for both J-aggregates and monomeric forms of JC-10 were measured with a FACSCalibur (Becton Dickinson) flow cytometer using FL1 and FL2 channels. Uncompensated data (left column) were compared with compensated data (right column).
Effect of FCCP induced mitochondria membrane potential change in Jurkat cells. Jurkat cells were dye loaded with JC-10 dye working solution along with DMSO (Top) or 5 µM FCCP (Low) for 10 minutes. The fluorescence intensities for both J-aggregates and monomeric forms of JC-10 were measured with a FACSCalibur (Becton Dickinson) flow cytometer using FL1 and FL2 channels. Uncompensated data (left column) were compared with compensated data (right column).
Ordering information
Price
Catalog Number
Unit Size
Quantity
Add to cart
Additional ordering information
Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
ShippingStandard overnight for United States, inquire for international
Request quotation
Spectral properties
Excitation (nm)508
Emission (nm)524
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Related products
Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Green Fluorescence*
Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Blue Fluorescence*
Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Blue Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Mitochondrial Hydroxyl Radical Detection Kit *Red Fluorescence*
Cell Meter™ Fluorimetric Mitochondrial Superoxide Activity Assay Kit *Green Fluorescence*
Cell Meter™ Fluorimetric Intracellular Peroxynitrite Assay Kit *Green Fluorescence*
Cell Meter™ Fluorimetric Intracellular Peroxynitrite Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *Orange Fluorescence Optimized for Microplate Reader*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *Orange Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Live Cell Caspase 3/7 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 3/7 Binding Assay Kit *Red Fluorescence*
Cell Meter™ Live Cell Caspase 1 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 2 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 6 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 8 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 9 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 10 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 13 Binding Assay Kit *Green Fluorescence*
Cell Meter™ No-Wash Live Cell Caspase 3/7 Activity Assay Kit *Blue Fluorescence*
Cell Meter™ No-Wash Live Cell Caspase 3/7 Activity Assay Kit *Red Fluorescence*
Cell Meter™ Fluorimetric Intracellular pH Assay Kit
Cell Meter™ Fluorimetric Cell Cytotoxicity Assay Kit
Cell Meter™ Cell Viability Assay Kit *Red Fluorescence*
Cell Meter™ Cell Viability Assay Kit *Blue Fluorescence with 405 nm Excitation*
Cell Meter™ Cell Viability Assay Kit *Blue Fluorescence*
Cell Meter™ Cell Viability Assay Kit *Green Fluorescence*
Cell Meter™ Cell Viability Assay Kit *NIR Fluorescence Optimized for Fluorescence Microplate Reader*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Blue Fluorescence Optimized for Microplate Readers*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Green Fluorescence Optimized for Microplate Readers*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Red Fluorescence Optimized for Microplate Readers*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Deep Red Fluorescence Optimized for Microplate Readers*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Orange Fluorescence Optimized for Microplate Readers*
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Blue Fluorescence*
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Caspase 8 Activity Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ NIR Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ NIR Mitochondrion Membrane Potential Assay Kit *Optimized for Microplate Reader*
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Orange Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Orange Fluorescence Optimized for Microplate Reader*
Cell Meter™ Intracellular GSH Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ Nuclear Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Caspase 8 Activity Apoptosis Assay Kit *Blue Fluorescence*
Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Blue Fluorescence*
Cell Meter™ Caspase 8 Activity Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Multiplexing Caspase 3/7, 8 and 9 Activity Assay Kit *Triple Fluorescence Colors*
Cell Meter™ Generic Fluorimetric Caspase Activity Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Generic Fluorimetric Caspase Binding Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Orange Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Deep Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Blue Fluorescence Excited at 405 nm*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Green Fluorescence Excited at 405 nm*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Orange Fluorescence Excited at 405 nm*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Deep Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Blue Fluorescence Excited at 405 nm*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Green Fluorescence Excited at 405 nm*
Cell Meter™ APC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ PE-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ Apoptotic and Necrotic Multiplexing Detection Kit I *Triple Fluorescence Colors*
Cell Meter™ Fluorimetric Live Cell Cycle Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Fixed Cell Cycle Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Apoptotic and Necrotic Multiplexing Detection Kit II *Triple Fluorescence Colors*
Cell Meter™ Live Cell TUNEL Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Fluorimetric Live Cell Cycle Assay Kit *Optimized for 405 nm Violet Laser Excitation*
Cell Meter™ Live Cell Caspase 3/7 and Phosphatidylserine Detection Kit *Triple Fluorescence Colors*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Green Fluorescence*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Red Fluorescence*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Orange Fluorescence*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Deep Red Fluorescence*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Mitochondrial Superoxide Activity Assay Kit*Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Mitochondrial Superoxide Activity Assay Kit*Optimized for Microplate Reader*
Cell Meter™ Autophagy Assay Kit *Blue Fluorescence*
Cell Meter™ Autophagy Fluorescence Imaging Kit
Cell Meter™ Autophagy Assay Kit *Green Fluorescence*
Cell Meter™ FITC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *NIR Fluorescence Optimized for Microplate Reader*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *NIR Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Colorimetric WST-8 Cell Quantification Kit
Cell Meter™ Multiplexing Live, Apoptotic and Necrotic Cell Detection Kit III *Triple Fluorescence Colors*
Cell Meter™ 2-NBDG Glucose Uptake Assay Kit
Cell Meter™ Intracellular NADH/NADPH Fluorescence Imaging Kit *Red Fluorescence*
Cell Meter™ Intracellular NADH/NADPH Flow Cytometric Analysis Kit *Red Fluorescence*
Cell Meter™ Live Cell TUNEL Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ No Wash and Probenecid-Free Endpoint Calcium Assay Kit *Optimized for microplate reader*
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Red Fluorescence Optimized for Microplate Reader*
Cell Meter™ Intracellular Colorimetric Lipid Peroxidation (MDA) Assay Kit
Cell Meter™ Flow Cytometric Calcium Assay Kit
Cell Meter™ Fluorimetric Live Cell Cycle Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Cellular Lipid Peroxidation Assay Kit
Cell Meter™ Intracellular NADH/NADPH Fluorescence Imaging Kit *Deep Red Fluorescence*
Cell Meter™ Intracellular NADH/NADPH Flow Cytometric Analysis Kit *Deep Red Fluorescence*
Cell Meter™ Intracellular GSH Assay Kit *Optimized for Flow Cytometry with 405 nm excitation*
Cell Meter™ Mitochondrial Autophagy Imaging Kit *Red Fluorescence*
Cell Meter™ BX520 fixable viability dye
Cell Meter™ BX590 fixable viability dye
Cell Meter™ BX650 fixable viability dye
Cell Meter™ IX830 fixable viability dye
Cell Meter™ RX660 fixable viability dye
Cell Meter™ RX700 fixable viability dye
Cell Meter™ RX780 fixable viability dye
Cell Meter™ VX450 fixable viability dye
Cell Meter™ VX500 fixable viability dye
Cell Meter™ Live Cell Caspase 8 Binding Assay Kit *Red Fluorescence*
Cell Meter™ Fixed Cell and Tissue TUNEL Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ Fixed Cell and Tissue TUNEL Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Fluorimetric Cellular Voltage Assay Kit
Cell Meter™ Cell Adhesion Assay Kit
Cell Meter™ Fixed Cell and Tissue TUNEL Apoptosis Assay Kit *Deep Red Fluorescence*
Cell Meter™ Fixed Cell and Tissue TUNEL Apoptosis Assay Kit *Blue Fluorescence*
Cell Meter™ Fluorescence Gap Junction Tracing Kit
Cell Meter™ Colorimetric MTT Cell Proliferation Kit
Cell Meter™ Live Cell Caspase 3/7 Imaging Kit *Green Fluorescence*
Cell Meter™ Beta-Arrestin Translocation GPCR Signaling Kit
Cell Meter™ Live Cell ATP Assay Kit
Cell Meter™ Colorimetric Cell Cytotoxicity Assay Kit
Cell Meter™ Fluorimetric Phagocytosis Assay Kit *Red Fluorescence*
Cell Meter™ Cellular Senescence Activity Assay Kit *Green Fluorescence*
Cell Meter™ Cellular Senescence Activity Assay Kit *Red Fluorescence*
Cell Meter™ Cell Proliferation Assay Kit
Cell Meter™ Glucose Uptake Imaging Kit *Red Fluorescence*
Cell Meter™ Colorimetric MTS Cell Proliferation Kit
Show More (121)

OverviewpdfSDSpdfProtocol


Excitation (nm)
508
Emission (nm)
524
Although JC-1 is widely used in many labs, its poor water solubility causes extraordinary inconvenience. Even at 1 µM concentration, JC-1 tends to precipitate in aqueous buffer. JC-10 is developed to be a superior alternative to JC-1 where high dye concentration is desired. Compared to JC-1, JC-10 has much better water solubility. JC-10 is capable of entering selectively into mitochondria, and changes reversibly its color from green to orange as membrane potentials increase. This property is due to the reversible formation of JC-10 aggregates upon membrane polarization that causes shifts in emitted light from 520 nm (i.e., emission of JC-10 monomeric form) to 570 nm (i.e., emission of J-aggregate). When excited at 490 nm, the color of JC-10 changes reversibly from green to greenish orange as the mitochondrial membrane becomes more polarized. Both colors can be detected using the filters commonly mounted in all flow cytometers, so that green emission can be analyzed in fluorescence channel 1 (FL1) and greenish orange emission in channel 2 (FL2). Besides its potential use in flow cytometry, it can also be used in fluorescence imaging and fluorescence microplate platform. This kit provides all the essential components with an optimized assay method for the detection of apoptosis in cells with the loss of mitochondrial membrane potential. This fluorometric assay is based on the detection of the mitochondrial membrane potential changes in cells by the cationic, lipophilic JC-10 dye. In normal cells, JC-10 concentrates in the mitochondrial matrix where it forms red fluorescent aggregates. However, in apoptotic and necrotic cells, JC-10 exists in monomeric form and stains cells in green fluorescence. The kit is optimized for screening of apoptosis activators and inhibitors by flow cytometry. We also offer a convenient 96-well and 384-well fluorescence microtiter-plate format kit (cat#22800) for high through put screening.

Platform


Flow cytometer

Excitation488 nm laser
Emission530/30 nm, 575/26 nm filter
Instrument specification(s)FITC, PE channel

Components


Example protocol


AT A GLANCE

Protocol summary

  1. Prepare cells with test compounds at the density of 5 × 105 to 1 x 106 cells/mL
  2. Resuspend the cells in 500 µL of JC-10 working solution (2-5 × 105 cells/tube) 
  3. Incubate at 37°C or room temperature for 15-60 minutes
  4. Analyze with flow cytometer using FL1 channel (green fluorescence monomeric signal) and FL2 channel (orange fluorescence aggregated signal)

Important notes
Thaw all the kit components at room temperature before starting the experiment.

PREPARATION OF WORKING SOLUTION

Add 25 µL of 200X JC-10 (Component A) into 5 mL of Assay Buffer A (Component B) and mix well to make JC-10 working solution. Protect from light.

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

SAMPLE EXPERIMENTAL PROTOCOL

  1. Treat cells with test compounds for a desired period of time to induce apoptosis. Set up parallel control experiments.

    For Negative Control: Treat cells with vehicle only.

    For Positive Control: Treat cells with FCCP or CCCP at 2-10  µM in a 37 oC, 5% CO2 incubator for 15 to 30 minutes. Note: CCCP or FCCP can be added simultaneously with JC-10 working solution. Titration of the CCCP or FCCP may be required for optimal results with an individual cell lines. 

  2. Centrifuge the cells to get 2 - 5 × 105 cells per tube. Note: For adherent cells, gently lift the cells by 0.5 mM EDTA to remain the cells intake, and wash the cells once with serum-containing media prior to incubation with JC-10 working solution.

  3. Resuspend cells in 500 µL of JC-10 working solution.

  4. Incubate the cells at room temperature or in a 37 oC, 5% CO2 incubator for 15 - 60 minutes, protected from light. Note: The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.

  5. Monitor the fluorescence intensity using flow cytometer with FL1 channel for the green fluorescence monomeric signal (in apoptotic cells), and FL2 channel for the orange fluorescence aggregated signal (in healthy cells). Gate on the cells, excluding debris. It is recommended that compensation corrections be performed using the FCCP or CCCP-treated cells.
  • Typical Flow Cytometer settings for the analysis of JC-10 on a BD FACS Calibur System flow cytometer are as follows:
    Suggested initial conditions may require modifications because of differences in cell types and culture conditions, and also the individual instrumentation.
    FL1 PMT voltage 366 
    FL2 PMT voltage 430
    Compensation: FL1 – 47.2% FL2; FL2 – 47.0% FL1

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)508
Emission (nm)524

Images


Citations


View all 86 citations: Citation Explorer
Search for new biologically active compounds: in vitro studies of antitumor and antimicrobial activity of dirhodium (II, II) paddlewheel complexes
Authors: Mitrovi{\'c}, Marina and Djuki{\'c}, Maja B and Vuki{\'c}, Milena and Nikoli{\'c}, Ivana and Radovanovi{\'c}, Marko D and Lukovi{\'c}, Jovan and Filipovi{\'c}, Ignjat P and Mati{\'c}, Sanja and Markovi{\'c}, Tijana and Klisuri{\'c}, Olivera R and others,
Journal: Dalton Transactions (2024)
Amorphous CaCO3-bioreactor for tumor microenvironment regulation to reinforce tumor chemoimmunotherapy
Authors: Chen, Weiguo and Lu, Yishuang and Xu, Yize and Chen, Yujiang and Lin, Shuai and He, Xin and Zhang, Chunfeng and Yuan, Chunsu
Journal: Chemical Engineering Journal (2024): 151838
Chemical diversity and antimelanoma potential of rosemary essential oils: Unveiling mechanistic insights through quantitative proteomics
Authors: Huang, Yeqin and Ding, Mengting and Wang, Di and Li, Hui and Xia, Fei and Bai, Hongtong and Sun, Meiyu and Mo, Meiling and Dong, Yanmei and Shi, Lei
Journal: Industrial Crops and Products (2024): 118652
Diabetes Primes Neutrophils for Neutrophil Extracellular Trap Formation through Trained Immunity
Authors: Shrestha, Sanjeeb and Lee, Yu-Bin and Lee, Hoyul and Choi, Yeon-Kyung and Park, Bo-Yoon and Kim, Mi-Jin and Youn, Young-Jin and Kim, Sun-Hwa and Jung, Soo-Jung and Song, Dong-Keun and others,
Journal: Research (2024)
TSPO deficiency exacerbates acute lung injury via NLRP3 inflammasome-mediated pyroptosis
Authors: Han, Jingyi and Zhang, Xue and Cai, Menghua and Tian, Feng and Xu, Yi and Chen, Hui and He, Wei and Zhang, Jianmin and Tian, Hui
Journal: Chinese Medical Journal (2024): 10--1097
Critical role of endoplasmic reticulum stress on bisphenol A-induced cytotoxicity in human keratinocyte HaCaT cells
Authors: Kim, Sou Hyun and Kang, Dong Wan and Kwon, Doyoung and Jung, Young-Suk
Journal: Environmental Toxicology (2024)
Altered lactate/pyruvate ratio may be responsible for aging-associated intestinal barrier dysfunction in male rats
Authors: Papila, Berrin and Karimova, Ayla and Onaran, Ilhan
Journal: Biogerontology (2024): 1--11
Mitochondria-synthesize melatonin to improve the quality and in vitro maturation as well as the embryonic development of porcine oocytes
Authors: Zhu, Tianqi and Wang, Likai and Ma, Xiao and Li, Guangdong and Fu, Yao and Ji, Pengyun and Zhang, Lu and Liu, Guoshi and others,
Journal: (2024)