Invitrogen™ MitoProbe™ JC-1 Assay Kit

The MitoProbe™ JC-1 Assay Kit is a quick and reliable assay designed to detect changes in mitochondria using flow cytometry. It leverages the JC-1 dye, a membrane-permeant dye widely used in apoptosis studies to monitor mitochondrial health. The JC-1 dye exhibits potential-dependent accumulation in mitochondria, which is indicated by a shift in fluorescence emission from green (∼529 nm) to red (∼590 nm). This shift makes the JC-1 assay a highly effective tool for studying mitochondrial membrane potential, which is critical for understanding early apoptosis and mitochondrial dysfunction.

The MitoProbe™ JC-1 Assay Kit includes a simple and streamlined protocol for compensated fluorescence ratio detection, allowing researchers to analyze the green-to-red fluorescence ratio and evaluate mitochondrial membrane potential changes in response to stimuli.

Key Features:

1. Mitochondrial Membrane Potential Indicator:
   • The JC-1 dye exhibits potential-dependent accumulation in mitochondria, providing a sensitive marker for monitoring changes in membrane potential in live cells.
2. Apoptosis Detection:
   • The shift from green to red fluorescence indicates changes in mitochondrial membrane potential, which is an early indicator of apoptosis. This makes the JC-1 assay particularly useful for early apoptosis detection in cell death studies.
3. Fluorescence Ratio Detection:
   • The green-to-red fluorescence ratio is used to quantify mitochondrial membrane potential changes, independent of other factors like mitochondrial size or density.
4. Compensated Fluorescence:
   • The kit includes CCCP (Mitochondrial Membrane Potential Disruptor), which helps to produce compensation controls for the green-to-red fluorescence ratio, ensuring accurate and reliable results.
5. Simple, Streamlined Protocol:
   • The assay protocol is easy to follow: incubate the cells with JC-1 dye and then analyze the fluorescence in the FITC and PE channels using a 488 nm laser on a flow cytometer.
6. Optimized for Flow Cytometry:
   • Specifically designed for use with flow cytometry systems, the JC-1 dye provides clear and consistent results when detecting mitochondrial health and apoptosis.
7. No PCR Requirement:
   • As the JC-1 dye is used in live cells, it is not dependent on PCR, providing a non-invasive method to monitor mitochondrial function.
8. Room Temperature Storage:
   • The kit is stored at room temperature and comes with vials of powdered dye, CCCP, and phosphate-buffered saline for easy use.

Specifications:

What's Included in the Kit:

• 5 vials of JC-1 dye (30 μg/vial of powdered dye)
• 1 vial of DMSO (1.5 mL)
• 1 vial of CCCP (125 μL, 50 mM in DMSO)
• 1 bottle of 10X phosphate-buffered saline (25 mL)

Applications:

1. Mitochondrial Health Monitoring:
   • Used to assess the mitochondrial membrane potential in live cells, helping to identify early signs of mitochondrial dysfunction and apoptosis.
2. Apoptosis Research:
   • Widely used in cell death studies to monitor changes in mitochondrial membrane potential as an early indicator of apoptosis.
3. Metabolic Studies:
   • Helps evaluate metabolic shifts and the impact of drugs or other treatments on mitochondrial function.
4. Cancer Research:
   • Useful for detecting apoptosis in cancer cells and evaluating how various treatments affect mitochondrial health.
5. Pharmaceutical Testing:
   • Employed to test the effects of pharmaceuticals on mitochondrial function and evaluate the impact on cell viability and cellular stress.

The Invitrogen™ JC-1 Assay Kit is a highly effective mitochondrial membrane potential probe that helps researchers assess mitochondrial health, detect apoptosis, and monitor cellular stress in live cells. Its ratiometric fluorescence mechanism makes it a reliable tool for mitochondrial studies, especially in cancer research, metabolic studies, and drug testing.