Molecular Probes™ MitoProbe™ JC-1 Assay Kit

Description

• Description The MitoProbe™ JC-1 Assay Kit is a quick and reliable assay used to detect changes in mitochondria by flow cytometry
• View a selection guide for all apoptosis assays for flow cytometry
• Selective to mitochondrial changes The membrane-permeant JC-1 dye is widely used in apoptosis studies to monitor mitochondrial health
• JC-1 dye exhibits potential-dependent accumulation in mitochondria, indicated by a fluorescence emission shift from green (∼529 nm) to red (∼590 nm)
• Consequently, mitochondrial depolarization is indicated by a decrease in the red/green fluorescence intensity ratio
• The potential-sensitive color shift is due to concentration-dependent formation of red fluorescent J-aggregates
• Unlike other mitochondrial stains that are affected by plasma membrane potential, the ratio of green to red fluorescence of JC-1 depends only on the membrane potential and not other factors such as mitochondrial size, shape, and density, which may influence single-component fluorescence signals
• Use of 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
• Indicator of early apoptosis A distinctive feature of the early stages of programmed cell death is the disruption of active mitochondria
• This mitochondrial disruption includes changes in the membrane potential and alterations to the oxidation-reduction potential of the mitochondria
• Changes in the membrane potential are presumed to be due to the opening of the mitochondrial permeability transition pore (MPTP), allowing passage of ions and small molecules
• The resulting equilibration of ions leads in turn to the decoupling of the respiratory chain and the release of cytochrome c into the cytosol
• Simple, streamlined protocol One of the biggest challenges to working with dual-emitting dyes like JC-1 is compensation
• The MitoProbe™ JC-1 Assay Kit provides a mitochondrial membrane-potential disrupter, CCCP, in order to produce compensation controls leading to a correctly compensated green-to-red fluorescence ratio
• The stain protocol is simple and straightforward: incubate the cells with JC-1 dye and then analyze the fluorescent signal in the FITC and PE channels of of a 488 nm laser.