Description
Key facts
Detection method:Fluorescent,
Sample types:Suspension cells, Adherent cells,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
How the assay works
JC-1 - mitochondrial membrane potential assay kit uses tetraethylbenzimidazolylcarbocyanine iodide (JC-1), a cationic dye that accumulates in energized mitochondria to measure the mitochondrial membrane potential. At low concentrations (due to low membrane potential) JC-1 is predominantly a monomer that yields green fluorescence with emission of 530±15 nm. At high concentrations (due to high membrane potential) the dye aggregates yielding a red to orange colored emission (590±17.5 nm). Therefore, a decrease in the aggregate fluorescent count is indicative of depolarization whereas an increase is indicative of hyperpolarization. The accompanying FCCP (carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone) is an ionophore uncoupler of oxidative phosphorylation. Treating cells with FCCP eliminates mitochondrial membrane potential and JC1 staining. JC1 is suitable for the labeling of mitochondria in live cells and it is not compatible with fixation.
JC-1 - Mitochondrial Membrane Potential Assay Kit protocol summary
- Wash cells in dilution buffer or PBS
- Add JC solution
- Incubate for 30 min at 37°C for suspension cells, or 10 min for adherent cells
- Wash cells with dilution buffer
- Treat cells as desired for experimental plan
- Analyze on a fluorescent microplate reader
The aggregate dye can be excited at 535 nm, the monomer and aggregate together at 475 nm.
JC-1 - Mitochondrial Membrane Potential Assay Kit has been used in a variety of sample type including:
Human fibroblasts and kidney cells
Human astrocytes
Human primary gingival fibroblasts
References:
1- Ziegler D et al. 2024
2-Mekhaeil M et al. 2023
3-Szymanska E et al. 2023
Related and recommended products
See other alternative kits to quantify Mitochondrial Membrane Potential:
- JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate)
ab112134
- JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry)
ab112133
- TMRE-Mitochondrial Membrane Potential Assay Kit
ab113852
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
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.
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.
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.
