MitoProbe™ JC-1 Assay Kit is a fast and reliable assay for detection by flow cytometry Mitochondrial changes.

ViewSelection guide for all apoptosis assays for flow cytometric analysis.

Selective for mitochondrial changes
The membrane-permeable JC-1 dye is widely used in apoptosis studies to monitor mitochondrial health. The fluorescence emission changes from green (∼529 nm) to red (∼590 nm), indicating that JC-1 dye accumulates in mitochondria and exhibits potential-dependent characteristics. Therefore, a decrease in the red/green fluorescence intensity ratio represents mitochondrial depolarization. The potential-sensitive color change is due to the concentration-dependent formation of red fluorescent J aggregates.

Unlike other mitochondrial stains that are affected by plasma membrane potential, the green-red fluorescence ratio of JC-1 only depends on the membrane potential and does not depend on other factors that may affect single-component fluorescence signals, such as mitochondria Size, shape and density. Using fluorescence ratio detection allows researchers to make comparative measurements of membrane potential and determine the percentage of mitochondria within a population that respond to an applied stimulus.

Indicators of early apoptosis
A special feature of early programmed cell death is active mitochondrial rupture. Mitochondrial rupture involves changes in membrane potential as well as changes in mitochondrial oxidation-reduction potential. The reason for the change in membrane potential may be due to the opening of the mitochondrial transition pore (MPTP), allowing ions and small molecules to enter the mitochondria. The resulting ionic balance results in the uncoupling of the respiratory chain and the release of cytochrome c into the cytosol.

Simple, simplified approach
One of the bigger challenges in using dual-emitting dyes such as JC-1 is compensation. The MitoProbe™ JC-1 Assay Kit provides the mitochondrial membrane potential disruptor CCCP to generate compensation control to obtain a correctly compensated green-to-red fluorescence ratio. The staining protocol is simple and straightforward: cells are incubated with JC-1 dye, and the fluorescence signal is analyzed using a 488 nm laser in the FITC and PE channels.