Mitochondrial membrane potential (ΔΨm) is a critical indicator of mitochondrial function and cellular health. Changes in ΔΨm are associated with early stages of apoptosis, mitochondrial dysfunction, and various disease states. Accurate and sensitive measurement of ΔΨm is essential for understanding cellular metabolism, drug responses, and toxicity mechanisms.

JC-1 is a sensitive, cationic dye widely used to detect changes in ΔΨm. JC-1’s dual-emission properties allow for ratiometric measurements, improving accuracy and reducing background interference. In healthy mitochondria with high membrane potential, JC-1 selectively accumulates and forms J-aggregates, which emit red fluorescence. In low potential or damaged mitochondria, JC-1 remains in its monomeric form, emitting green fluorescence. The shift from red to green fluorescence serves as a sensitive indicator of mitochondrial depolarization.

The RayBio^® JC-1 Mitochondrial Membrane Potential Assay Kit is an in vitro fluorescent assay designed for use with live cells. The assay enables detection and quantification of mitochondrial membrane potential changes via the red/green fluorescence intensity ratio, providing a reliable assessment of mitochondrial health under a broad range of experimental conditions. It is compatible with fluorescent microplate readers and fluorescent microscopes.

• Accurate and sensitive
• Compatible with fluorescence microscopy
• Suitable for plate-based readers

• Fluorescent plate reader or microscope capable of reading
  • Ex/Em: 525/590 nm (Red J-aggregate)
  • Ex/Em: 490/530 nm (Green monomer)

• General tissue culture supplies
• FBS (Fetal Bovine Serum)
• If using a microplate reader, sterile 96-well clear bottom, dark sided microplates with lids
• If using a microscope, sterile 8-well chamber slides w/ removable wells or 96-well microplates and microscope slides
• Adjustable pipettes (1–1000 μL)

Contact our technical support team for questions about compatibility: (techsupport@raybiotech.com)

1. Thaw and mix JC-1 and FCCP reagents.
2. Dilute JC-1 in culture medium.
3. Seed cells in a 96-well plate.
4. Add FCCP to positive control wells.
5. Remove supernatants from wells.
6. Add JC-1 staining solution to wells.
7. Incubate cells with JC-1 staining solution.
8. Measure fluorescence using a microplate reader or fluorescent microscope.