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Distinguish high and low mitochondrial membrane potential in living cells using JC-1 (Iodide) dye. JC-1 offers a fast and convenient staining protocol that is well suited for high-throughput screening and compatible with flow cytometry applications.
Features:
• Excitation wavelength: 515 nm
• Emission wavelength: 530 nm
• Extinction coefficient: 195,000 cm⁻¹M⁻¹
• Fluorescence based on dye concentration
• Verified for use with flow cytometry
JC-1 is a lipophilic, cationic carbocyanine dye. This dye can reversibly change its fluorescence, which depends on the equilibrium between its monomeric and aggregate form. JC-1 exists as a monomer in polarized mitochondria and, upon excitation, it emits a green fluorescence with an emission maximum of 530 nm. At higher concentrations, due to high mitochondrial membrane potential, J-aggregates form and emit a green-orange fluorescence with an emission maximum of 590 nm. JC-1 can be used both qualitatively to measure a shift in fluorescence emission, or quantitatively to detect the difference in fluorescence intensity in channels FL1 (FITC) and FL2 (PE).
The green to orange fluorescent intensity ratio was used to evaluate mitochondrial depolarization occurring in apoptosis (Sivandzade et al. Bio-protocol, 2019). JC-1 has been used to simultaneously evaluate drug resistance parameters P-glycoprotein (P-gp) activity and apoptosis in myeloid leukemia cells (Chaoui et al. Cytometry B Clin Cytom, 2006).
Figure 1. Mitochondrial Depolarization Is Visualized by a Shift to JC-1 (Iodide) Monomers
Flow cytometry analysis of Jurkat cells labeled with JC-1 (Iodide) after incubation in the presence or absence of staurosporine, an inducer of apoptosis. Mitochondrial depolarization is visualized by the population shift from JC-1 aggregates to JC-1 monomers after treatment with staurosporine.
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