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JC-1

5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide; CAS#: 3520-43-2
JC-1 is a green-orange fluorescent dye used to assay mitochondrial membrane potential for studying cell health in flow cytometry applications.

Mechanism and principle

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. In response to increases in mitochondrial transmembrane potential (ΔΨM), 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 ΔΨM), 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.

FormMitochondria Membrane PotentialColorAbsorption (nm)Emission (nm)Emission Channel
JC-1 monomerLowGreen510527FL1
J aggregateHighOrange585590FL2

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.

Analysis of the Mitochondrial Membrane Potential Using the Cationic JC-1 Dye as a Sensitive Fluorescent Probe

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 mg0.5 mg1 mg5 mg10 mg
1 mM153.32 µL766.601 µL1.533 mL7.666 mL15.332 mL
5 mM30.664 µL153.32 µL306.64 µL1.533 mL3.066 mL
10 mM15.332 µL76.66 µL153.32 µL766.601 µL1.533 mL

Molarity calculator

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

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum

Citations

View all 34 citations: Citation Explorer
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
Autophagy-deficient macrophages exacerbate cisplatin-induced mitochondrial dysfunction and kidney injury via miR-195a-5p-SIRT3 axis
Authors: Yuan, Yujia and Yuan, Longhui and Yang, Jingchao and Liu, Fei and Liu, Shuyun and Li, Lan and Liao, Guangneng and Tang, Xi and Cheng, Jingqiu and Liu, Jingping and others,
Journal: Nature Communications (2024): 4383
Palmitic Acid Exerts Anti-Tumorigenic Activities by Modulating Cellular Stress and Lipid Droplet Formation in Endometrial Cancer
Authors: Zhao, Ziyi and Wang, Jiandong and Kong, Weimin and Newton, Meredith A and Burkett, Wesley C and Sun, Wenchuan and Buckingham, Lindsey and O’Donnell, Jillian and Suo, Hongyan and Deng, Boer and others,
Journal: Biomolecules (2024): 601
Ipatasertib, an oral AKT inhibitor, in combination with carboplatin exhibits anti-proliferative effects in uterine serous carcinoma
Authors: Burkett, Wesley C and Zhao, Ziyi and Newton, Meredith A and Sun, Wenchuan and Deng, Boer and Secord, Angeles Alvarez and Zhou, Chunxiao and Bae-Jump, Victoria
Journal: Annals of Medicine (2023): 603--614
Page updated on September 27, 2024

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Physical properties

Molecular weight

652.23

Solvent

DMSO

Spectral properties

Extinction coefficient (cm -1 M -1)

1950001

Excitation (nm)

515

Emission (nm)

530

Storage, safety and handling

Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200

CAS

3520-43-2

Platform

Flow cytometer

Excitation488 nm laser
Emission530, 30 nm, 575, 26 nm filter
Instrument specification(s)FITC, PE channel

Fluorescence microscope

Excitation490 nm
Emission525 nm (590 nm for ratio analysis)
Recommended plateBlack wall, clear bottom
Instrument specification(s)FITC and TRITC filters

Fluorescence microplate reader

Excitation490 nm
Emission525 nm (590 nm for ratio analysis)
Recommended plateSolid black
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