JC‑10 is an ideal fluorescent probe for detecting mitochondrial membrane potential (ΔΨm). When the mitochondrial membrane potential is high, JC‑10 aggregates in the mitochondrial matrix and emits red fluorescence; when the membrane potential is low, JC‑10 exists as monomers and emits green fluorescence. JC‑10 selectively enters mitochondria and undergoes a reversible fluorescence color shift from green to orange‑red as the membrane potential increases. Compared with JC‑1, JC‑10 exhibits better water solubility while maintaining similar fluorescence properties, making it a superior alternative to JC‑1.
Product Use
For research use only. Widely applied in cell biology, molecular biology, pharmacology, and other research fields. Not for human use. JC‑10 is suitable for detecting membrane potential changes in cells, tissues, or isolated mitochondria.
Physical Properties and Specifications
Appearance: Liquid
Molecular Formula: C₂₅H₂₉Cl₂IN₄
Molecular Weight: 583.34
CAS No. 5563‑28‑0
Concentration: 2 mg/mL (equivalent to 3 mM)
Solubility: Clear in DMSO, free from visible impurities
Excitation/Emission Maxima (nm): 490/525 (monomer, green); 490/590 (aggregate, orange‑red)
Storage Conditions: Store at ‑20°C, protected from light and moisture, in airtight containers
Operating Protocol (For Reference Only)
1. Preparation of Working Solution
Thaw JC-10 solution to room temperature. Dilute it with HBSS (Hanks' Balanced Salt Solution with HEPES) or other buffer supplemented with 0.02% Pluronic F-127 to a final concentration of 5–30 μM, then vortex thoroughly for homogenization.
2. Sample Experimental Protocols
2.1 Microplate Reader Detection
(1) Treat cells with test compounds for scheduled duration to induce apoptosis (Example: Jurkat cells can be treated with camptothecin for 4–6 hours). Set up negative control wells (cells plus solvent control) and background blank wells (medium only without cells).
(2) Remove culture medium from adherent cells before staining; pellet suspension cells by centrifugation to discard medium prior to staining.
(3) Add 100 μL JC-10 working solution per well for 96-well plates, or 25 μL per well for 384-well plates.
(4) Incubate the plates in a 37 ℃ incubator with 5% CO₂ protected from light for 15–60 min.
Note: Incubation time should be optimized according to cell type and concentration; it is recommended to confirm the optimal incubation duration before each experiment.
Note: Incubation time should be optimized according to cell type and concentration; it is recommended to confirm the optimal incubation duration before each experiment.
Optional: After aspirating JC-10 working solution, add 100 μL HHBS buffer per well for 96-well plates (25 μL per well for 384-well plates) before detection and analysis.
(5) Monitor fluorescence signals at excitation/emission wavelengths of 490/525 nm and 490/590 nm for ratiometric analysis.
2.2 Detection via Fluorescence Microscope or Flow Cytometer
(1) Treat cells with test compounds for scheduled duration to induce apoptosis (Example: Jurkat cells can be treated with camptothecin for 4–6 hours).
(2) Centrifuge cells and adjust the cell density to 1–5×10⁵ cells per tube.
(3) Resuspend cell pellets in 500 μL JC-10 working solution.
(4) Incubate the cell suspension protected from light at room temperature or 37 ℃ for 10–60 min.
Note: Incubation time should be optimized according to cell type and concentration; it is recommended to confirm the optimal incubation duration before each experiment.
Optional: Remove JC-10 working solution by centrifugation, resuspend cells with an appropriate volume of HHBS buffer, then proceed to detection and analysis.
(5) Detect fluorescence changes under excitation/emission wavelengths of 490/525 nm and 490/590 nm using a fluorescence microscope equipped with FITC/TRITC filter sets, or a flow cytometer with FL1/FL2 channels.
Precautions
Fluorescent dyes are susceptible to photobleaching. Keep samples away from light to slow down fluorescence quenching.
For research use only by qualified professionals. Not for clinical diagnosis or therapy, food or pharmaceutical applications. Do not store in residential areas.
Wear a lab coat and disposable gloves during operation for your personal safety and health.
We only provide partial technical information for some products and do not guarantee the authenticity and authority of the data. All data above are for reference and academic research only.
