Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>

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).
Ordering information
Price ()
Catalog Number22801
Unit Size
Find Distributor
Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Spectral properties
Excitation (nm)508
Emission (nm)524
Storage, safety and handling
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
Show More (129)

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


Component A: 200X JC-10 in DMSO1 vial (250 µL)
Component B: Assay Buffer1 bottle (50 mL)

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

Citations


View all 58 citations: Citation Explorer
Impact of Organelle Transport Deficits on Mitophagy and Autophagy in Niemann--Pick Disease Type C
Authors: Liedtke, Maik and V{\"o}lkner, Christin and Hermann, Andreas and Frech, Moritz J
Journal: Cells (2022): 507
Mitochondria Transfer from Adipose Stem Cells Improves the Developmental Potential of Cryopreserved Oocytes
Authors: Kankanam Gamage, Udayanga Sanath and Hashimoto, Shu and Miyamoto, Yuki and Nakano, Tatsuya and Yamanaka, Masaya and Koike, Akiko and Satoh, Manabu and Morimoto, Yoshiharu
Journal: Biomolecules (2022): 1008
Patchouli alcohol suppresses castration-resistant prostate cancer progression by inhibiting NF-$\kappa$B signal pathways
Authors: Cai, Jian and Zhao, Juan and Gao, Ping and Xia, Yuguo
Journal: Translational Andrology and Urology (2022): 528
A comparative study of metformin and nicotinamide riboside in alleviating tissue aging in rats
Authors: Geng, Lingling and Zhang, Bin and Liu, Haisong and Wang, Si and Cai, Yusheng and Yang, Kuan and Zou, Zhiran and Jiang, Xiaoyu and Liu, Zunpeng and Li, Wei and others,
Journal: Life Medicine (2022)
DNA repair proteins as the targets for paroxetine to induce cytotoxicity in gastric cancer cell AGS
Authors: Liu, Bang-Hung and Yuan, Tein-Ming and Huang, Chih-Jou and Hsu, Duan-Ting and Chen, Shi-Wen and Hsiao, Nai-Wan and Lin, Sheng-Chih and Wu, Shu-Wan and Lin, Yi-Mei J and Chuang, Show-Mei
Journal: American journal of cancer research (2022): 1465
Effects of time-restricted feeding and type of food on fertility competence in female mice
Authors: Konishi, Nafuko and Matsumoto, Hiroshi and Hashimoto, Shu and Gamage, Udayanga Sanath Kankanam and Tachibana, Daisuke and Fukuda, Aisaku and Morimoto, Yoshiharu and Koyama, Masayasu
Journal: Scientific Reports (2022): 1--15
The relationship between free fatty acids and mitochondrial oxidative stress damage to trophoblast cell in preeclampsia
Authors: Jiang, Lingling and Yan, Jianying
Journal: BMC Pregnancy and Childbirth (2022): 1--6
Gracillin Isolated from Reineckia carnea Induces Apoptosis of A549 Cells via the Mitochondrial Pathway
Authors: Yang, Jianqiong and Cao, Ling and Li, Yamei and Liu, Hai and Zhang, Minhong and Ma, Huamou and Wang, Biao and Yuan, Xiaoliang and Liu, Qian
Journal: Drug design, development and therapy (2021): 233
Mitochondrial Function and Root-Filled Teeth--Detrimental and Unknown Interfaces in Systemic Immune Diseases
Authors: Lechner, Johann and Mayer, Wolfgang
Journal: International Journal of General Medicine (2020): 387--402
Inhibition of phosphodiesterase-10A by Papaverine protects human cortical neurons from quinolinic acid induced oxidative stress and synaptic proteins alterations
Authors: Bhat, Abid and Tan, Vanessa and Heng, Benjamin and Essa, Musthafa M and Chidambaram, Saravana B and Guillemin, Gilles J
Journal: bioRxiv (2020)