Invitrogen™ MitoProbe™ DiOC2(3) Assay Kit - For Flow Cytometry - 100 Assays

The MitoProbe™ DiOC2(3) Assay Kit is a fluorescence-based mitochondrial membrane potential assay designed for use in flow cytometry applications. The kit contains DiOC2(3), a membrane-potential-sensitive cyanine dye, and CCCP (Carbonyl cyanide 3-chlorophenylhydrazone), a mitochondrial depolarization agent for use as a positive control.

DiOC2(3) is a lipophilic cationic dye that penetrates the cytosol of live eukaryotic cells and accumulates in mitochondria in response to their membrane potential (ΔΨm). It fluoresces red in mitochondria with intact membrane potential and shifts to green fluorescence upon mitochondrial depolarization. This shift in fluorescence intensity allows for quantitative assessment of mitochondrial health using flow cytometry.

✔ Enables real-time detection of mitochondrial membrane potential (ΔΨm)

✔ Flow cytometry-compatible for high-throughput, single-cell analysis

✔ Red-to-green fluorescence shift for easy detection of mitochondrial depolarization

✔ Includes CCCP as a mitochondrial membrane potential disruptor for assay validation

✔ For Research Use Only. Not for diagnostic procedures.

Key Features & Benefits

✔ Quantitative Detection of Mitochondrial Function

• DiOC2(3) localizes in mitochondria and fluoresces red when the membrane potential is intact.
• Depolarized mitochondria exhibit a red-to-green fluorescence shift, providing a reliable indication of mitochondrial dysfunction.

✔ Optimized for Flow Cytometry Applications

• High-throughput single-cell analysis with flow cytometry.
• Detects shifts in mitochondrial membrane potential in live cells.
• Requires no washing steps, minimizing sample loss and variability.

✔ Mitochondrial Membrane Potential Disruptor (CCCP) Included

• CCCP serves as a positive control, ensuring accurate assessment of mitochondrial depolarization.
• Prevents dye accumulation by collapsing mitochondrial membrane potential.

✔ Rapid and Simple Protocol

• Minimal hands-on time; mix, incubate, and analyze by flow cytometry.
• No washing steps required, reducing the risk of cell loss or experimental variability.

✔ Ideal for Multiple Research Applications

• Apoptosis and cell viability studies
• Mitochondrial function assessment in neurodegeneration and cancer research
• Toxicology and drug screening assays
• Metabolic and bioenergetics studies

Applications & Use Cases

1. Mitochondrial Function and Bioenergetics Studies

• Purpose: Evaluate mitochondrial membrane potential as an indicator of mitochondrial function.
• Use Case: Researchers use this assay to assess mitochondrial activity in metabolic diseases, aging, and cellular energy metabolism research.

2. Apoptosis and Cell Death Analysis

• Purpose: Detect early mitochondrial depolarization in apoptotic cells.
• Use Case: Used in cancer research and pharmacology to evaluate the effect of drugs that induce apoptosis via mitochondrial pathways.

3. Toxicology and Drug Screening

• Purpose: Measure mitochondrial toxicity of pharmaceutical compounds.
• Use Case: Used in toxicology studies to assess the impact of drug candidates on mitochondrial function in primary cells and cell lines.

4. Neurodegeneration and Neurological Disorders Research

• Purpose: Investigate mitochondrial dysfunction associated with neurodegenerative diseases.
• Use Case: Used in Alzheimer’s, Parkinson’s, and ALS research to study mitochondrial involvement in neuronal cell death.

5. Cancer and Stem Cell Research

• Purpose: Analyze mitochondrial bioenergetics in tumor cells and stem cells.
• Use Case: Used in tumor biology and regenerative medicine to compare mitochondrial activity between normal and transformed cells.

Kit Specifications & Technical Details

Kit Contents & Storage

✔ Kit Includes:

• DiOC2(3) Staining Dye: 625 µL (10 µM in DMSO)
• CCCP Mitochondrial Membrane Potential Disruptor: 125 µL (50 mM in DMSO)

✔ Storage Conditions:

• Store at 2–6°C (protected from light).
• Do not freeze, as freezing can degrade the reagents.

How the Assay Works

Step-by-Step Protocol:

1. Prepare Cell Samples
   • Suspend cells in an appropriate buffer or culture medium.
2. Stain with DiOC2(3) Dye
   • Add DiOC2(3) directly to live cells in a flow cytometry-compatible format.
3. Incubate the Sample
   • Allow the dye to accumulate in mitochondria with an intact membrane potential.
4. Add CCCP (Optional, for Control Experiments)
   • If assessing mitochondrial depolarization, treat a portion of cells with CCCP to induce mitochondrial membrane potential loss.
5. Analyze by Flow Cytometry
   • Measure fluorescence intensity shifts from red to green in mitochondria using a flow cytometer.

Ordering Information

Regulatory & Compliance Information

• For Research Use Only (RUO) – Not intended for diagnostic procedures.
• Manufactured under strict quality control standards for high reproducibility and performance.

The MitoProbe™ DiOC2(3) Assay Kit is a powerful and reliable flow cytometry tool for assessing mitochondrial membrane potential changes in live cells. Its red-to-green fluorescence shift allows researchers to quantify mitochondrial depolarization, making it ideal for apoptosis studies, toxicology testing, neurodegenerative disease research, and cancer metabolism investigations.

With its easy-to-use workflow, high specificity, and compatibility with flow cytometry, the MitoProbe™ DiOC2(3) Assay Kit is an essential tool for mitochondrial function analysis in life science research.