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.
Fluorescent
Suspension cells, Adherent cells
Direct
20m
Select an associated product type
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.
Fluorescent
Suspension cells, Adherent cells
Direct
20m
Flow cytometer
Blue Ice
-20°C
-20°C
-20°C
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 μM 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 .**Related assays**Review the to learn about kits to perform a , and . Review the to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
This supplementary information is collated from multiple sources and compiled automatically.
The mitochondrial membrane potential also known as ΔΨm is the electrical potential difference across the inner mitochondrial membrane. This potential results from the electrochemical gradient produced by the proton pumps during electron transport chain activity. The mechanical function of the mitochondrial membrane potential is important to ATP production through oxidative phosphorylation. Mitochondrial membranes are widely expressed in almost all eukaryotic cells and are an essential component of cellular metabolism. The inner membrane is structured to facilitate its function housing integral proteins that are key to maintaining the potential.
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.
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.
Changes in the mitochondrial membrane potential relate significantly to conditions like neurodegenerative diseases and cancer. In neurodegenerative diseases such as Parkinson’s and Alzheimer's dysregulation of mitochondrial membrane potential can lead to impaired energy production and increased oxidative stress. Cancer cells often exhibit altered mitochondrial membrane potential affecting processes like apoptosis and enabling survival in adverse conditions. These alterations in potential impact proteins such as p53 which play critical roles in cancer progression and neurodegenerative disease pathology.
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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).
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