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Chemical Structure| 3520-43-2 Chemical Structure| 3520-43-2

Structure of JC-1
CAS No.: 3520-43-2

Chemical Structure| 3520-43-2

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JC-1 (CBIC2) is an ideal fluorescent probe widely used to detect mitochondrial membrane potential. JC-1 accumulates in mitochondria in a potential-dependent manner and is used to detect membrane potential in cells, tissues, or purified mitochondria. In normal mitochondria, JC-1 aggregates in the mitochondrial matrix to form a polymer that emits strong red fluorescence (Ex=585nm, Em=590nm). When the mitochondrial membrane potential is low, JC-1 cannot aggregate and produces green fluorescence (Ex=514nm, Em=529nm).

Synonyms: CBIC2; NK 1420

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Product Citations

Product Citations

Lockhart, Steven ; Babu, Dinesh ; Tran, Newton H ; Reiz, Béla ; Tonoyan, Lusine ; Siraki, Arno G

Abstract: 6-PPD (N-[1,3-dimethylbutyl]-N′-phenyl-p-phenylenediamine) is an industrial antioxidant reported to be an environmental contaminant. It was found to be highly toxic to coho salmon and potentially other aquatic organisms. The toxicity of 6-PPD in humans, however, remains unknown. The neutrophil enzyme myeloperoxidase (MPO) is known to catalyze xenobiotic metabolism; therefore, its role in 6-PPD cytotoxicity was investigated using the MPO-rich HL-60 cell line. UV−visible spectroscopy and liquid chromatography−mass spectrometry (LC/MS) were performed to investigate the MPO-mediated oxidation of 6-PPD and identify possible metabolites in the absence and presence of (GSH). 6-PPD’s cytotoxicity, effect on mitochondrial membrane potential (MMP), and GSH-depleting ability in HL-60 cells were assessed. Electron paramagnetic resonance (EPR) was used to determine radical formation using DMPO, and mitochondrial-derived superoxide was assessed with the mito-TEMPO-H probe. Evaluation of the 6- PPD-induced cellular injury pathways was performed by preincubating an antioxidant and an MPO inhibitor with HL-60 cells. UV−vis analysis of MPO-catalyzed oxidation of 6-PPD demonstrated changes in the 6-PPD spectrum, whereas the addition of altered the spectrum, indicating possible conjugate formation. LC/MS showed the formation of multiple products, including GSH-6-PPD conjugates and a conjugate to a (a known 6-PPD degradant), which could potentially induce cytotoxicity. 6-PPD demonstrated concentration-dependent cytotoxicity, and cellular levels were decreased by 6-PPD. Similarly, the level of MMP decreased, suggesting mitochondrial depolarization. Furthermore, the EPR spin probe for mitochondrial superoxide showed a positive relationship with 6-PPD concentration, and EPR spin-trapping demonstrated 6-PPD concentration-dependent radical signal intensity using MPO/H2O2. The precursor, NAC, demonstrated partial cytoprotection against 6-PPD; however, the MPO inhibitor surprisingly showed no significant cytoprotective effect. Our results suggest that MPO could be a potential catalyst for 6-PPD toxicity in humans. However, MPO inhibition did not significantly affect cellular viability, suggesting an MPO-independent toxicity pathway. These findings warrant a deeper investigation to determine 6-PPD mammalian toxicity pathways.

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Product Details of JC-1

CAS No. :3520-43-2
Formula : C25H27Cl4IN4
M.W : 652.23
SMILES Code : CC[N+]1=C(/C=C/C=C2N(CC)C3=CC(Cl)=C(Cl)C=C3N2CC)N(CC)C4=CC(Cl)=C(Cl)C=C41.[I-]
Synonyms :
CBIC2; NK 1420
MDL No. :MFCD00188475

Safety of JC-1

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302-H315-H319
Precautionary Statements:P501-P270-P264-P280-P302+P352-P337+P313-P305+P351+P338-P362+P364-P332+P313-P301+P312+P330

Isoform Comparison

Biological Activity

Description
JC-1 (CBIC2) is an ideal fluorescent probe for detecting mitochondrial membrane potential, accumulating in mitochondria in a potential-dependent manner. It can be used to assess the membrane potential of cells, tissues, or purified mitochondria. In healthy mitochondria, JC-1 aggregates in the mitochondrial matrix, forming a polymer that emits strong red fluorescence; when mitochondrial membrane potential decreases, JC-1 cannot aggregate, producing green fluorescence instead[1].

In Vitro:

Cell Line
Concentration Treated Time Description References
cardiomyocytes 5 μM 10 min To evaluate mitochondrial membrane potential, results showed that doxorubicin treatment significantly reduced mitochondrial membrane potential, and MIF deficiency exacerbated this effect. PMC3886760
H9C2 cells 1 μM 24 h To assess the effect of IF1 knockdown on mitochondrial membrane potential in H9C2 cells, results showed that IF1 knockdown cells had a less pronounced isoproterenol-induced ΔΨm reduction compared to scramble shRNA H9C2 cells PMC5585346
HK-2 cells 10 µg/mL 15 min JC-1 was used to assess mitochondrial membrane potential (MMP), showing that the MMP was significantly reduced in WT mice after ischemia-reperfusion (I/R), while the MMP reduction was less pronounced in RIP3-/- mice. PMC9220894
SH-SY5Y cells 1 μM 24 h JC-1 staining was used to examine changes in mitochondrial membrane potential. In the MPP+-induced SH-SY5Y cell model, the JC-1 red fluorescence intensity declined to 55.48% and green fluorescence intensity was increased. After treatment with compounds 63, 68, and 74, the red MFIs of JC-1 were markedly recovered to 77.23%, 75.46%, and 68.61%, respectively. PMC9928544
Primary neurons 1 μM 24 h JC-1 staining was used to examine changes in mitochondrial membrane potential. Compounds 63, 68, and 74 significantly alleviated the decrease in mitochondrial membrane potential caused by MPP+ and significantly reduced cellular oxidative damage. PMC9928544

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
ICR male mice Photoaging mouse model Gavage 50 mg/kg and 200 mg/kg Once daily for 10 weeks To evaluate the effects of JC and JCH on skin moisture, histological changes, and immunity indexes in photoaging mice. The results showed that JC and JCH could repair endogenous collagen and elastin protein fibers, maintain the natural ratio of type I to type III collagen, and enhance the immunity of photoaging mice. PMC3571645

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

1.53mL

0.31mL

0.15mL

7.67mL

1.53mL

0.77mL

15.33mL

3.07mL

1.53mL

Dissolving Methods
Please choose the appropriate dissolution scheme according to your animal administration guide.For the following dissolution schemes, clear stock solution should be prepared according to in vitro experiments, and then cosolvent should be added in turn:

in order to ensure the reliability of the experimental results, the clarified stock solution can be properly preserved according to the storage conditions; The working fluid for in vivo experiment is recommended to be prepared now and used on the same day;

The percentage shown in front of the following solvent refers to the volume ratio of the solvent in the final solution; If precipitation or precipitation occurs in the preparation process, it can be assisted by heating and/or ultrasound.
Protocol 1
Protocol 2

References

 

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