Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Microplate Assays*
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Spectral properties
Excitation (nm) | 508 |
Emission (nm) | 524 |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Alternative formats
Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry Assays* |
Related products
Overview | SDSProtocol |
See also: Mitochondria, Mitochondrial Membrane Potential
Excitation (nm) 508 | Emission (nm) 524 |
Although JC-1 is widely used in many labs, its poor water solubility makes it hard to use for some applications. Even at 1 µM concentration, JC-1 tends to precipitate in aqueous buffer. JC-10 has been developed to be a superior alternative to JC-1 where high dye concentration is desired. Compared to JC-1, our 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. This Cell Meter™ JC-10 Mitochondrial Membrane Potential Assay Kit enable you to monitor mitochondrial membrane potential changes using a simple microplate reader while all the other commercial JC-1 assay kits require the use of a flow cytometer. Our kit provides the most robust method to monitor mitochondrial membrane potential changes, and can be readily used for screening a large compound library.
Platform
Fluorescence microplate reader
Excitation | 490/540 nm |
Emission | 525/590 nm |
Cutoff | 515/570 nm |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Bottom read mode |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add test compounds
- Add JC-10 dye-working solution (50 µL/well/96-well plate or 12.5 µL/well/384-well plate)
- Incubate at 37°C, 5% CO2 incubator for 30 to 60 minutes
- Add Assay Buffer B (50 µL/well/96-well plate or 12.5 µL/well/384-well plate)
- Monitor fluorescence intensities (bottom read mode) at Ex/Em = 490/525 nm (Cutoff = 515 nm) and 540/590 nm (Cutoff = 570 nm)
Important notes
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
Add 50 µL of 100X JC-10 (Component A) into 5 mL of Assay Buffer A (Component B) and mix well to make JC-10 dye-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
- Treat cells by adding 10 µL of 10X test compounds (96-well plate) or 5 µL of 5X test compounds (384-plate) into the desired buffer (such as PBS or HHBS). Note: It is not necessary to wash cells before adding compound. However, if tested compounds are serum sensitive, growth medium and serum factors can be aspirated away before adding compounds. Add the same volume of HHBS into the wells (such as 90 µL for a 96-well plate or 20 µL for a 384-well plate) after aspiration. Alternatively, cells can be grown in serum-free media.
- Incubate the cell plate at room temperature or in a 37°C, 5% CO2 incubator for at least 15 minutes or a desired period of time (for Jurkat cells, 4 - 6 hours with camptothecin or 3 - 5 hours with staurosporine treatment) to induce apoptosis.
- Add 50 µL/well (96-well plate) or 12.5 µL/well (384-well plate) of JC-10 dye-working solution into the cell plate.
- Incubate the plate in a 37°C, 5% CO2 incubator for 30 - 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.
- Add 50 µL/well (96-well plate) or 12.5 µL/well (384-well plate) of Assay Buffer B (Component C) into JC-10 dye-working solution plate before reading the fluorescence intensity. Note: DO NOT wash the cells after loading. For non-adherent cells, it is recommended to centrifuge cell plates at 800 rpm for 2 minutes with brake off after adding Assay Buffer B (Component C).
- Monitor the fluorescence intensities with a fluorescence microplate reader (bottom read mode) at Ex/Em = 490/525 nm (Cutoff = 515 nm) and 540/590 nm (Cutoff = 570 nm) for ratio analysis.
Images
Figure 1. Campotothecin-induced mitochondria membrane potential changes were measured with JC-10™ and JC-1 in Jurkat cells. After Jurkat cells were treated with camptothecin (10 µM) for 4 hours, JC-1 and JC-10™ dye working solutions were added to the wells and incubated for 30 minutes. The fluorescence intensities for both J-aggregates (highlighted in blue) and monomeric forms (highlighted in red) of JC-1 and JC-10™ were measured at Ex/Em = 490/525 nm (Cutoff = 515 nm) and 490/590 nm (Cutoff = 570 nm) with NOVOstar microplate reader (BMG Labtech).
Citations
View all 76 citations: Citation Explorer
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Journal: Journal of Controlled Release (2024): 694--711
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Journal: Journal of Controlled Release (2024): 694--711
Ginsenosides Inhibit the Proliferation of Lung Cancer Cells and Suppress the Rate of Metastasis by Modulating EGFR/VEGF Signaling Pathways
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Journal: Journal of Oleo Science (2024): 219--230
Authors: Yu, Xuelian and Wang, Qihu and Dai, Zhaoxin
Journal: Journal of Oleo Science (2024): 219--230
Mitochondria-synthesize melatonin to improve the quality and in vitro maturation as well as the embryonic development of porcine oocytes
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The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progression
Authors: Wang, Ying and Wu, Zhimin and Yan, Guoqiang and Li, Shan and Zhang, Yanzhuo and Li, Guangping and Wu, Chengai
Journal: Bone \& Joint Research (2024): 4
Authors: Wang, Ying and Wu, Zhimin and Yan, Guoqiang and Li, Shan and Zhang, Yanzhuo and Li, Guangping and Wu, Chengai
Journal: Bone \& Joint Research (2024): 4
Bushen Huoxue Formula Inhibits IL-1$\beta$-Induced Apoptosis and Extracellular Matrix Degradation in the Nucleus Pulposus Cells and Improves Intervertebral Disc Degeneration in Rats
Authors: Gao, Shang and Wang, Chenmoji and Qi, Lijie and Liang, Songlin and Qu, Xintian and Liu, Wei and Li, Nianhu
Journal: Journal of Inflammation Research (2024): 121--136
Authors: Gao, Shang and Wang, Chenmoji and Qi, Lijie and Liang, Songlin and Qu, Xintian and Liu, Wei and Li, Nianhu
Journal: Journal of Inflammation Research (2024): 121--136
Mitochondrial dysfunction induced by bedaquiline as an anti-Toxoplasma alternative
Authors: Shi, Yuehong and Jiang, Yucong and Qiu, Haolong and Hu, Dandan and Song, Xingju
Journal: Veterinary Research (2023): 1--16
Authors: Shi, Yuehong and Jiang, Yucong and Qiu, Haolong and Hu, Dandan and Song, Xingju
Journal: Veterinary Research (2023): 1--16
Non-apoptotic regulated cell death in Fuchs endothelial corneal dystrophy
Authors: Sakakura, Saki and Inagaki, Emi and Sayano, Tomoko and Yamazaki, Risa and Fusaki, Noemi and Hatou, Shin and Hirayama, Masatoshi and Tsubota, Kazuo and Negishi, Kazuno and Okano, Hideyuki and others,
Journal: Regenerative Therapy (2023): 592--601
Authors: Sakakura, Saki and Inagaki, Emi and Sayano, Tomoko and Yamazaki, Risa and Fusaki, Noemi and Hatou, Shin and Hirayama, Masatoshi and Tsubota, Kazuo and Negishi, Kazuno and Okano, Hideyuki and others,
Journal: Regenerative Therapy (2023): 592--601
The role of mitochondria in nanoparticle-specific stem cell toxicity
Authors: WITTERS, Margo and SAENEN, Nelly and VAN BELLEGHEM, Frank and BRONCKAERS, Annelies and SMEETS, Karen
Journal: (2023)
Authors: WITTERS, Margo and SAENEN, Nelly and VAN BELLEGHEM, Frank and BRONCKAERS, Annelies and SMEETS, Karen
Journal: (2023)
(E)-2-Octenal suppresses the growth of a prochloraz-resistant Penicillium italicum strain and its potential antifungal mechanisms
Authors: Luo, Yuwei and Chen, Xin and Che, Jin Xin and Zhang, Yonghua and Ouyang, Qiuli and Tao, Nengguo
Journal: Postharvest Biology and Technology (2023): 112515
Authors: Luo, Yuwei and Chen, Xin and Che, Jin Xin and Zhang, Yonghua and Ouyang, Qiuli and Tao, Nengguo
Journal: Postharvest Biology and Technology (2023): 112515
Hydroxyhydroquinone and Quassinoids as Promising Compounds with Hypoglycemic Activity through Redox Balance
Authors: dos Santos, Paulo Roberto and Danetti, Sidineia and Rastegar, Joseph and De Souza, Wellington and Frassini, Rafaele and Scariot, Fernando Joel and Moura, Sidnei and Roesch-Ely, Mariana
Journal: (2023)
Authors: dos Santos, Paulo Roberto and Danetti, Sidineia and Rastegar, Joseph and De Souza, Wellington and Frassini, Rafaele and Scariot, Fernando Joel and Moura, Sidnei and Roesch-Ely, Mariana
Journal: (2023)
Application notes
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
Fluorescent Oligonucleotide Labeling Reagents
Monitoring of Mitochondrial Membrane Potential Changes in Live Cells Using JC-10
Selective Analysis of RNA in Live and Fixed Cells with StrandBrite RNA Green
Cell Loading Protocol For Fluorescent pH Indicator, BCECF-AM
Fluorescent Oligonucleotide Labeling Reagents
Monitoring of Mitochondrial Membrane Potential Changes in Live Cells Using JC-10
Selective Analysis of RNA in Live and Fixed Cells with StrandBrite RNA Green
Cell Loading Protocol For Fluorescent pH Indicator, BCECF-AM
FAQ
Does propidium iodide stain apoptotic cells?
I ordered your phalloidin-amine (Cat #5302) so I can conjugate it to my oligo. Do you have a recommended protocol I can use?
What dye works best for staining and tracking lysosomes in live cells for several hours?
How can I lyse my cells without lysing the nuclear membrane?
Do you have any dual-fluorescence nucleic acid stains that interact with both DNA and RNA?
I ordered your phalloidin-amine (Cat #5302) so I can conjugate it to my oligo. Do you have a recommended protocol I can use?
What dye works best for staining and tracking lysosomes in live cells for several hours?
How can I lyse my cells without lysing the nuclear membrane?
Do you have any dual-fluorescence nucleic acid stains that interact with both DNA and RNA?