JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry)
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Flow Cytometry - JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (AB112133)
JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133) was used to measure the effect of FCCP induced mitochondria membrane potential change in Jurkat cells by Flow Cytometry. Jurkat cells were dye loaded with JC-10 dye-loading solution along with DMSO (Top) or 5 μM FCCP (Low) for 10 minutes. The fluorescent intensities for both J-aggregates and monomeric forms of JC-10 were measured with a flow cytometer using FL1 and FL2 channels. Uncompensated data (left column) were compared with compensated data (right column).
Product details
JC-10 Mitochondrial Membrane Potential Assay Kit ab112133 is designed for use with flow cytometry, and it provides the most robust assay method for monitoring changes in mitochondrial membrane potential.
The 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 diffuses out of mitochondria, changes to a monomeric form and stains cells with green fluorescence.
Although JC-1 is widely used in many labs, its poor water solubility causes great inconvenience. Even at 1 uM concentration, JC-1 tends to precipitate in aqueous buffer. Compared to JC-1, JC-10 has much better water solubility.
JC-10 selectively enters mitochondria, and reversibly changes its color from green to orange-red as membrane potentials increase. This property is due to the reversible formation of JC-10 aggregates upon membrane polarization which cause a shifts in emitted light from 520 nm (the emission of JC-10 monomeric form) to 570 nm (the emission of JC-10-aggregate form). When excited at 490 nm, the color of JC-10 changes reversibly from green to greenish orange as the mitochondrial membrane becomes more polarized.
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 green. The green emission can be analyzed in fluorescence channel 1 (FL1) and greenish orange emission in channel 2 (FL2). Both colors can be detected using the filters commonly mounted in all flow cytometers. Besides its use in flow cytometry, it can also be used in fluorescence imaging and fluorescence microplate platform.
JC-10 assay protocol summary:
- add JC-10 staining solution to experimentally treated cells
- incubate cells for 15-60 min
- analyze wth flow cytometer
If you would like to use JC-10 on a microplate reader, we recommend ab112134.
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Biological function summary
The mitochondrial membrane potential drives ATP synthesis by powering ATP synthase an enzyme complex embedded in the mitochondrial membrane. This potential also plays a vital role in other processes such as calcium homeostasis and regulation of mitochondrial biogenesis. The mitochondrial membrane itself forms part of the larger mitochondrial respiratory chain complex coordinating with components like complex I (NADH: ubiquinone oxidoreductase) and complex II (succinate dehydrogenase) to maintain cell energy needs and respond to metabolic demands.
Pathways
The mitochondrial membrane potential is integral to cellular energy metabolism pathways such as the Krebs cycle and oxidative phosphorylation. Mitochondrial membrane potential modulation can affect signaling proteins like cytochrome c which is instrumental in apoptosis. Apoptotic signaling pathways involving proteins such as Bax and Bcl-2 influence the mitochondrial membrane potential and regulate cell survival or death in response to cellular stress or damage.
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Publications (14)
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Veterinary research communications 49:214 PubMed40471485
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Microcirculation (New York, N.Y. : 1994) 32:e70012 PubMed40394906
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PloS one 20:e0322733 PubMed40315213
2025
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Journal of biomedical science 30:63 PubMed37537557
2023
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iScience 26:107446 PubMed37599822
2023
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Biomolecules 12: PubMed35740913
2022
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Frontiers in immunology 12:753683 PubMed34899705
2021
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Cancer genomics & proteomics 18:645-659 PubMed34479917
2021
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Anticancer research 41:2963-2977 PubMed34083287
2021
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NPJ Regenerative medicine 6:28 PubMed34039998
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