JC-1 [5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide] *CAS#: 3520-43-2*
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See also: Mitochondria, Cell Metabolism, Flow Cytometry Reagents, Cell Structures and Organelles, Membrane Potential and Channels
CAS 3520-43-2 | Molecular weight 652.23 | Extinction coefficient (cm -1 M -1) 1950001 | Excitation (nm) 515 | Emission (nm) 530 |
JC-1 is widely used for determining mitochondrial membrane potential with flow cytometry. It is capable of entering selectively into mitochondria, and changes reversibly its color from green to orange as membrane potentials increase (over values of about 80-100 mV). This property is due to the reversible formation of JC-1 aggregates upon membrane polarization that causes shifts in emitted light from 530 nm (i.e., emission of JC-1 monomeric form) to 590 nm (i.e., emission of J-aggregate). When excited at 490 nm, the color of JC-1 changes reversibly from green to greenish orange as the mitochondrial membrane becomes more polarized. Both colors can be detected using the filters commonly mounted in all flow cytometers, so that green emission can be analyzed in fluorescence channel 1 (FL1) and greenish orange emission in channel 2 (FL2). The main advantage of the use of JC-1 is that it can be both qualitative, considering the shift from green to orange fluorescence emission, and quantitative, considering the pure fluorescence intensity, which can be detected in both FL1 and FL2 channels. Besides its wide use with flow cytometry, it is also used in fluorescence imaging. We have developed a protocol to use it in fluorescence microplate platform. Although JC-1 is widely used in many labs, its poor water solubility makes it hard to use for some applications. Our JC-10 has much better water solubility than JC-1, and in some cell lines JC-10 has even superior performance to JC-1. Interestingly the performance of JC-10 is quite cell line-dependent.
Platform
Flow cytometer
Excitation | 488 nm laser |
Emission | 530/30 nm, 575/26 nm filter |
Instrument specification(s) | FITC, PE channel |
Fluorescence microscope
Excitation | 490 nm |
Emission | 525 nm (590 nm for ratio analysis) |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | FITC and TRITC filters |
Fluorescence microplate reader
Excitation | 490 nm |
Emission | 525 nm (590 nm for ratio analysis) |
Recommended plate | Solid black |
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
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Spectrum
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Spectral properties
Extinction coefficient (cm -1 M -1) | 1950001 |
Excitation (nm) | 515 |
Emission (nm) | 530 |
Citations
View all 26 citations: Citation Explorer
Mitochondrial transfer from MSCs to macrophages restricts inflammation and alleviates kidney injury in diabetic nephropathy mice via PGC-1$\alpha$ activation
Authors: Yuan, Yujia and Yuan, Longhui and Li, Lan and Liu, Fei and Liu, Jingping and Chen, Younan and Cheng, Jingqiu and Lu, Yanrong
Journal: STEM CELLS (2021)
Authors: Yuan, Yujia and Yuan, Longhui and Li, Lan and Liu, Fei and Liu, Jingping and Chen, Younan and Cheng, Jingqiu and Lu, Yanrong
Journal: STEM CELLS (2021)
Ruxolitinib induces apoptosis of human colorectal cancer cells by downregulating the JAK1/2-STAT1-Mcl-1 axis
Authors: Li, Xia and Wang, Zhe and Zhang, Shengjie and Yao, Qinghua and Chen, Wei and Liu, Feiyan
Journal: Oncology letters (2021): 1--14
Authors: Li, Xia and Wang, Zhe and Zhang, Shengjie and Yao, Qinghua and Chen, Wei and Liu, Feiyan
Journal: Oncology letters (2021): 1--14
Pogostemon cablin Triggered ROS-Induced DNA damage to arrest cell cycle progression and induce apoptosis on human hepatocellular carcinoma in vitro and in vivo
Authors: Huang, Xiao-Fan and Sheu, Gwo-Tarng and Chang, Kai-Fu and Huang, Ya-Chih and Hung, Pei-Hsiu and Tsai, Nu-Man
Journal: Molecules (2020): 5639
Authors: Huang, Xiao-Fan and Sheu, Gwo-Tarng and Chang, Kai-Fu and Huang, Ya-Chih and Hung, Pei-Hsiu and Tsai, Nu-Man
Journal: Molecules (2020): 5639
Cotargeting the JAK/STAT signaling pathway and histone deacetylase by ruxolitinib and vorinostat elicits synergistic effects against myeloproliferative neoplasms
Authors: Hao, Xing and Xing, Wen and Yuan, Jiajia and Wang, Yingshao and Bai, Jiaojiao and Bai, Jie and Zhou, Yuan
Journal: Investigational new drugs (2019): 1--11
Authors: Hao, Xing and Xing, Wen and Yuan, Jiajia and Wang, Yingshao and Bai, Jiaojiao and Bai, Jie and Zhou, Yuan
Journal: Investigational new drugs (2019): 1--11
Mitochondrial ROS-induced lysosomal dysfunction impairs autophagic flux and contributes to M1 macrophage polarization in a diabetic condition
Authors: Yuan, Yujia and Chen, Younan and Peng, Tianqing and Li, Lan and Zhu, Wuzheng and Liu, Fei and Liu, Shuyun and An, Xingxing and Luo, Ruixi and Cheng, Jingqiu and others, undefined
Journal: Clinical Science (2019): 1759--1777
Authors: Yuan, Yujia and Chen, Younan and Peng, Tianqing and Li, Lan and Zhu, Wuzheng and Liu, Fei and Liu, Shuyun and An, Xingxing and Luo, Ruixi and Cheng, Jingqiu and others, undefined
Journal: Clinical Science (2019): 1759--1777
Inhibition der PI3K/AKT/mTOR-Signaltransduktion: innovative Therapieoptionen f{\"u}r neuroendokrine Tumore auf Basis eines tieferen Verst{\"a}ndnisses intrazellul{\"a}rer Prozesse in NEN
Authors: Freitag, Helma
Journal: (2018)
Authors: Freitag, Helma
Journal: (2018)
overexpression of protocadherin 7 inhibits neuronal survival by downregulating Birc5 in vitro
Authors: Xiao, Huajuan and Sun, Ziling and Wan, Jun and Hou, Shengtao and Xiong, Yi
Journal: Experimental cell research (2018): 71--80
Authors: Xiao, Huajuan and Sun, Ziling and Wan, Jun and Hou, Shengtao and Xiong, Yi
Journal: Experimental cell research (2018): 71--80
Therapeutic potential of GSK-J4, a histone demethylase KDM6B/JMJD3 inhibitor, for acute myeloid leukemia
Authors: Li, Yunan and Zhang, Mingying and Sheng, Mengyao and Zhang, Peng and Chen, Zizhen and Xing, Wen and Bai, Jie and Cheng, Tao and Yang, Feng-Chun and Zhou, Yuan
Journal: Journal of Cancer Research and Clinical Oncology (2018): 1--13
Authors: Li, Yunan and Zhang, Mingying and Sheng, Mengyao and Zhang, Peng and Chen, Zizhen and Xing, Wen and Bai, Jie and Cheng, Tao and Yang, Feng-Chun and Zhou, Yuan
Journal: Journal of Cancer Research and Clinical Oncology (2018): 1--13
Cell death mechanisms of the anti-cancer drug etoposide on human cardiomyocytes isolated from pluripotent stem cells
Authors: Nemade, Harshal and Chaudhari, Umesh and Acharya, Aviseka and Hescheler, Jürgen and Hengstler, Jan Georg and Papadopoulos, Symeon and Sachinidis, Agapios
Journal: Archives of Toxicology (2018): 1--18
Authors: Nemade, Harshal and Chaudhari, Umesh and Acharya, Aviseka and Hescheler, Jürgen and Hengstler, Jan Georg and Papadopoulos, Symeon and Sachinidis, Agapios
Journal: Archives of Toxicology (2018): 1--18
Charakterisierung der Apoptoseinduktion im Rhabdomyosarkommodell: Analyse eines triterpens{\"a}urehaltigen Viscum album Gesamtextraktes
Authors: Stammer, Rahel Mascha
Journal: (2018)
Authors: Stammer, Rahel Mascha
Journal: (2018)
Application notes
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Monitoring of Mitochondrial Membrane Potential Changes in Live Cells Using JC-10
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
Activation of the Mitochondrial Apoptotic Pathway Produces Reactive Oxygen Species and Oxidative Damage in Hepatocytes That Contribute to Liver Tumorigenesis
Calmodulin antagonists induce cell cycle arrest and apoptosis in vitro and inhibit tumor growth in vivo in human multiple myeloma