JC-1 - Mitochondrial Membrane Potential Assay Kit
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(2 Publications)
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Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
GM14643 healthy leukocytes were treated with 0, 25, 50, or 100uM FCCP in complete media for 30 minutes. Cells were collected by centrifugation and washed once with serum free media. Cells were resuspended with a staining solution of 5uM or 10uM JC-1 in serum free media and incubated for 30 minutes at 37°C in the dark. Cells were collected by centrifugation, washed once with 1XPBS, and resuspended in 1X Opti-Klear. Cells were then seeded in a 96-well optical bottom plate at a density of 200,000 cells/50uL/well. Plate was imaged using the CX5 CellInsight.
- FuncS
Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
HeLa cells plated at 12000 cells/well. Next day, aspirated media and replaced with complete media containing 0 or 100uM FCCP for 30 minutes. Drug solution aspirated from wells and cells were washed once with serum free media. A staining solution of 1uM JC-1 in serum free media was added to cells and incubated for 15 minutes at 37°C in the dark. Staining solution was aspirated, cells were washed once with 1X PBS, and bathed in 1X Opti-Klear. Plate was imaged using the CX5 CellInsight.
- FuncS
Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
JC-1 assay result in HL60 cells treated with Troglitazone. HL60 cells were seeded and labeled according to section 11.1 of the protocol. Cells were then treated for 4 hours with a titration series of the thiazolidinedione Troglitazone and both monomer and aggregate forms were read on a Perkin Elmer-Wallac 1420 Victor 2 Multilabel plate reader. Mean and standard deviation of aggregate/monomer ratios is plotted for 3 replicates for each concentration. IC50 of Troglitazone in HL60 cells was calculated at 1.2 μM
- FuncS
Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
GM14643 healthy leukocytes were treated with 0, 25, 50, or 100uM FCCP in complete media for 30 minutes. Cells were collected by centrifugation and washed once with serum free media. Cells were resuspended with a staining solution of 5uM or 10uM JC-1 in serum free media and incubated for 30 minutes at 37°C in the dark. Cells were collected by centrifugation, washed once with 1XPBS, and resuspended in 1X Opti-Klear. Cells were then seeded in a 96-well optical bottom plate at a density of 200,000 cells/50uL/well. Fluorescence was measured on Tecan plate reader using Ex. 485/Em. 530 for monomer and Ex. 535/Em. 590 for aggregates.
- FuncS
Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
HeLa cells plated at 12000 cells/well. Next day, aspirated media and replaced with complete media containing 0, 25, 50, or 100uM FCCP for 30 minutes. Drug solution aspirated from wells and cells were washed once with serum free media. A staining solution of 1uM JC-1 in serum free media was added to cells and incubated for 15 minutes at 37°C in the dark. Staining solution was aspirated, cells were washed once with 1X PBS, and bathed in 1X Opti-Klear. Plate was analyzed using the CX5 CellInsight SpotDetector bioapp
- FuncS
Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
HeLa cells plated at 12000 cells/well. Next day, aspirated media and replaced with complete media containing 0, 25, 50, or 100uM FCCP for 30 minutes. Drug solution aspirated from wells and cells were washed once with serum free media. A staining solution of 1uM JC-1 in serum free media was added to cells and incubated for 15 minutes at 37°C in the dark. Staining solution was aspirated, cells were washed once with 1X PBS, and bathed in 1X Opti-klear. Measured fluorescence on Tecan plate reader using Ex. 485/Em. 530 for monomer and Ex. 535/Em. 590 for aggregates.
- FuncS
Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
GM14643 healthy leukocytes were treated with 0, 25, 50, or 100uM FCCP in complete media for 30 minutes. Cells were collected by centrifugation and washed once with serum free media. Cells were resuspended with a staining solution of 5uM or 10uM JC-1 in serum free media and incubated for 30 minutes at 37°C in the dark. Cells were collected by centrifugation, washed once with 1XPBS, and resuspended in 1X Opti-Klear. Cells were then seeded in a 96-well optical bottom plate at a density of 200,000 cells/50uL/well. Plate was analyzed using the CX5 CellInsight SpotDetector bioapp
- FuncS
Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
JC-1 assay result in HL60 cells treated with FCCP. HL60 cells were seeded and labeled according to section 11.1 of the protocol. Cells were then treated for 4 hours with 100 μM FCCP or vehicle/diluent control (DMSO). Cells were read on a Perkin Elmer-Wallac 1420 Victor 2 Multilabel plate reader. Mean and standard deviation is plotted for 3 replicates from each condition.
- FuncS
Supplier Data
Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB288313)
JC1 - Mitochondrial Membrane Potential Assay Kit.
JC-1 assay result in HepG2 cells treated with CCCP. HepG2 cells were seeded and labeled according to section 11.2 of the protocol. Cells were then treated for 4 hours with a titration series of CCCP (carbonyl cyanide 3-chlorophenylhydrazone) and both monomer and aggregate forms were read on a Perkin Elmer-Wallac 1420 Victor 2 Multilabel plate reader. Mean and standard deviation of aggregate/monomer ratios is plotted for 12 replicates for each concentration. IC50 of CCCP in HepG2 cells was calculated at 8.7 μM
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Product details
JC1- Mitochondrial Membrane Potential Assay Kit ab288313 contains tetraethylbenzimidazolylcarbocyanine iodide (JC-1), a cationic dye that accumulates in energized mitochondria.
At low concentrations (due to low mitochondrial membrane potential), JC-1 is predominantly a monomer that yields green fluorescence with emission of 530±15 nm.
At high concentrations (due to high mitochondrial 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 JC-1 staining protocol is very simple:
- 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.
Review our to learn more about our other , and assay kits. 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.
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Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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 (2)
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FASEB bioAdvances 7:e70009 PubMed40330431
2025
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Journal of neurochemistry 162:290-304 PubMed35598091
2022
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