JC-1 Mitochondrial Membrane Potential Dye

JC-1

5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide; CAS#: 3520-43-2

JC-1 is a mitochondrial membrane potential indicator that exhibits green fluorescence as monomers and red fluorescence as J-aggregates, providing ratiometric assessment of mitochondrial health and apoptosis.

Mitochondrial Membrane Potential Sensor: Green monomers (525 nm) shift to red aggregates (590 nm) at high ΔΨm
Ratiometric Analysis: Red/green ratio quantifies mitochondrial polarization state
Lipophilic Cation: Accumulates in mitochondria proportional to membrane potential
Flow Cytometry Compatible: Dual-color detection for single-cell bioenergetics analysis

Chemical structure for JC-1 [5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide] *CAS#: 3520-43-2*

Protocol

COO

COA

Catalog	Size	Price	Quantity
22200	5 mg	Price
22201	50 mg	Price

Overview

JC-1 (5,5,6,6’-tetrachloro-1,1’,3,3’ tetraethylbenzimi-dazoylcarbocyanine iodide) is a cationic dye that selectively labels mitochondria in live cells to assay mitochondrial membrane potential in flow cytometry applications. In response to increases in mitochondrial transmembrane potential (ΔΨ_M), from approximately 80 to 100 mV, the dye changes color from green to orange. The mechanism for this is the reversible formation of JC-1 aggregates upon membrane polarization; high membrane potential results in JC-1 monomer aggregation into J aggregates. These J aggregates exhibit orange fluorescence, while JC-1 monomers fluoresce green. By monitoring the intensity and ratios of green/orange fluorescences, JC-1 staining can be used as an assay for mitochondrial membrane potential.

Excitation and emission

Under low mitochondrial membrane potential conditions, JC-1 molecules exist largely as monomers. These have an absorption peak of 510 nm and an emission peak of 527 nm. Under polarized conditions (high ΔΨ_M), J aggregates form, which have an absorption peak of 585 nm and an emission peak of 590 nm. Both JC-1 monomers (green form) and J aggregates (orange form) can be excited by a 488 nm argon laser. The JC-1 monomer green emission can be detected using the FL1 channel of a flow cytometer, while the J aggregate orange emission can be detected using the FL2 channel.

Form	Mitochondria Membrane Potential	Color	Absorption (nm)	Emission (nm)	Emission Channel
JC-1 monomer	Low	Green	510	527	FL1
J aggregate	High	Orange	585	590	FL2

JC-1 Alternatives

Although JC-1 is widely used in many labs, one problem it suffers from is poor water solubility. An alternative version, which addresses this issue, is JC-10.

Sivandzade, F., Bhalerao, A., & Cucullo, L. (2019). Analysis of the Mitochondrial Membrane Potential Using the Cationic JC-1 Dye as a Sensitive Fluorescent Probe. Bio-protocol, 9(1), e3128. https://doi.org/10.21769/BioProtoc.3128

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of JC-1 [5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide] *CAS#: 3520-43-2* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	153.32 µL	766.601 µL	1.533 mL	7.666 mL	15.332 mL
5 mM	30.664 µL	153.32 µL	306.64 µL	1.533 mL	3.066 mL
10 mM	15.332 µL	76.66 µL	153.32 µL	766.601 µL	1.533 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	652.23 g/mol	=	mL	x	mM	=	mmol

Citations

View all 37 citations: Citation Explorer

MT2A Promotes Angiogenesis in Chronically Ischemic Brains through a Copper--Mitochondria Regulatory Mechanism

Authors:

Mo, Ni and Tai, Chuyang and Yang, Yang and Ling, Cong and Zhang, Baoyu and Wei, Lei and Wang, Hui and Chen, Chuan and others,

Lithocholic Acid’s Ionic Compounds as Promising Antitumor Agents: Synthesis and Evaluation of the Production of Reactive Oxygen Species (ROS) in Mitochondria

Authors:

Chobanov, Nuri M and Dzhemileva, Lilya U and Dzhemilev, Usein M and D’yakonov, Vladimir A

Journal:

Antioxidants (2024): 1448

Self-Assembled Nanocarriers of Synthetic and Natural Plasmalogens for Potential Nanomedicine Development

Authors:

Wu, Yu and Angelov, Borislav and Deng, Yuru and Fujino, Takehiko and Hossain, Md Shamim and Bizien, Thomas and Angelova, Angelina

Journal:

Advanced Therapeutics (2024): 2400093

Rhopaloic acid A triggers mitochondria damage-induced apoptosis in oral cancer by JNK/BNIP3/Nix-mediated mitophagy

Authors:

Chen, Wu-Fu and Tsai, Sheng-Chieh and Zhang, Ya-Hui and Chang, Hui-Min and Wu, Wan-Ju and Su, Jui-Hsin and Wu, Bin-Nan and Chen, Chung-Yi and Lin, Mei-Ying and Chen, Hsien-Lin and others,

Journal:

Phytomedicine (2024): 155855

Insulin reduces endoplasmic reticulum stress-induced apoptosis by decreasing mitochondrial hyperpolarization and caspase-12 in INS-1 pancreatic $\beta$-cells

Authors:

Murata, Nanako and Nishimura, Kana and Harada, Naoki and Kitakaze, Tomoya and Yoshihara, Eiji and Inui, Hiroshi and Yamaji, Ryoichi

Journal:

Physiological Reports (2024): e16106

Physical properties

Molecular weight	652.23
Solvent	DMSO

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	195000 ¹
Excitation (nm)	515
Emission (nm)	530

Storage, safety and handling

Certificate of Origin	Download PDF
H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200
CAS	3520-43-2

Instrument settings

Flow cytometer
Excitation	488 nm laser
Emission	530/30 nm, 575/26 nm filter
Instrument specification(s)	FITC and PE channel

Fluorescence microscope
Excitation	FITC and TRITC filter set
Emission	FITC and TRITC filter set
Recommended plate	Black wall/clear bottom

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Page updated on October 13, 2025