3,3′-Diethyloxacarbocyanine Iodide 97%

3,3′-Diethyloxacarbocyanine iodide (DiOC2(3)) is a fluorescent carbocyanine dye primarily used for measuring mitochondrial membrane potential (ΔΨm) in live cells. As a cationic, lipophilic probe, it accumulates in mitochondria proportional to membrane potential.

Primary applications include:

• Apoptosis detection: Early indicator of mitochondrial depolarization
• Cell viability: Assessment in flow cytometry and microscopy
• Mitochondrial function: Real-time monitoring of metabolic activity
• Drug screening: Toxicity assessment and efficacy studies
• Flow cytometry: Multi-parameter cell analysis
• Live-cell imaging: Mitochondrial dynamics and distribution

DiOC2(3) is particularly valuable because mitochondrial depolarization is an early event in apoptosis, occurring before nuclear changes and cell membrane damage.

DiOC2(3) has the following spectral properties:

Spectral characteristics:

• Excitation maximum: 482 nm
• Emission maximum: 497 nm
• Extinction coefficient: ~150,000 M⁻¹cm⁻¹
• Stokes shift: ~15 nm

Compatible filter sets:

• FITC filter set (most common)
• Flow cytometry: FL1 channel (488 nm laser excitation)
• Microscopy: Standard FITC/GFP filter cubes

The green fluorescence makes DiOC2(3) compatible with most standard fluorescence equipment and allows for multi-color applications with red-emitting dyes like propidium iodide (PI) or Texas Red.

ChemContract supplies 3,3′-Diethyloxacarbocyanine iodide at ≥97% purity determined by HPLC analysis, suitable for all research applications.

Quality control testing:

• HPLC purity: ≥97% with chromatogram
• UV-Vis spectroscopy: λmax verification at 482 nm
• ¹H NMR: Structure confirmation
• Mass spectrometry: Molecular weight verification
• Functional testing: Available upon request

Certificate of Analysis (COA) includes:

• Batch number and manufacturing date
• HPLC chromatogram with purity determination
• UV-Vis spectrum with λmax values
• ¹H NMR spectrum
• Appearance and solubility data
• Storage recommendations

DiOC2(3) should be prepared as a concentrated stock solution in DMSO for optimal stability and solubility.

Stock solution preparation:

• Recommended concentration: 1-10 mM in anhydrous DMSO
• Dissolve completely: Vortex or sonicate briefly
• Protect from light: Wrap vial in aluminum foil
• Aliquot for storage: Divide into single-use aliquots
• Storage: -20°C in amber vials or foil-wrapped tubes
• Stability: 6-12 months when stored properly

Working solution preparation:

• Flow cytometry: Dilute stock to 20-40 nM in cell culture medium or PBS
• Microscopy: Dilute stock to 1-10 nM in appropriate buffer
• Plate reader: Dilute stock to 10-100 nM
• DMSO concentration: Keep final DMSO <0.1% for cell viability

Important notes:

• Always protect solutions from light during preparation and use
• Use immediately after dilution for best results
• Avoid repeated freeze-thaw cycles of stock solutions

Standard staining protocol for DiOC2(3) is straightforward and works for most cell types.

Basic staining protocol:

• Step 1: Prepare cells in PBS or culture medium (10⁶ cells/mL)
• Step 2: Add DiOC2(3) to final concentration of 20-40 nM (flow cytometry) or 1-10 nM (microscopy)
• Step 3: Incubate 15-30 minutes at 37°C, protected from light
• Step 4: Optional: Wash once with PBS or buffer
• Step 5: Analyze immediately by flow cytometry or microscopy

Important considerations:

• Temperature: 37°C incubation provides best results
• Incubation time: 15-30 minutes is sufficient for most cells
• Light protection: Keep samples in dark during and after staining
• Concentration optimization: May need adjustment for different cell types
• Controls: Include unstained and positive/negative controls

For apoptosis studies:

• Combine with propidium iodide (PI) to distinguish viable, apoptotic, and necrotic cells
• Use positive control (e.g., CCCP, valinomycin) to induce depolarization
• DiOC2(3)-low = depolarized mitochondria (early apoptosis)
• DiOC2(3)-high = polarized mitochondria (viable cells)

Proper storage is critical for maintaining DiOC2(3) stability and fluorescence properties.

Storage conditions:

• Temperature: -20°C (freezer storage)
• Light protection: Store in amber vials or wrap in aluminum foil
• Desiccation: Keep in desiccator to prevent moisture exposure
• Container: Tightly sealed to minimize air exposure
• Shelf life: 24 months when stored properly

Stock solution storage:

• Aliquoting: Divide into single-use aliquots to avoid freeze-thaw
• Temperature: -20°C in amber vials
• Stability: 6-12 months in DMSO
• Thawing: Warm to room temperature before use

Handling precautions:

• Minimize exposure to light during weighing and preparation
• Use powder immediately after weighing or return to freezer
• Avoid repeated freeze-thaw cycles
• Do not store diluted working solutions

Signs of degradation:

• Color change from dark green/black
• Reduced fluorescence intensity
• Precipitate formation in DMSO stock

We offer DiOC2(3) in multiple pack sizes for immediate delivery, with custom quantities available.

Stock quantities (ships 24-48 hours):

• 10 mg – Ideal for initial studies (~22 µmol)
• 25 mg – Standard research quantity (~56 µmol)
• 50 mg – Extended studies (~112 µmol)
• 100 mg – Long-term projects (~225 µmol)

Custom quantities:

• Bulk orders: 250 mg, 500 mg, 1g+ available
• Lead time: 1-2 weeks for custom quantities
• Volume pricing: Discounts available for large orders

Usage estimate:

• 10 mg makes ~22 mL of 1 mM stock (sufficient for ~200-400 flow cytometry samples)
• 25 mg makes ~56 mL of 1 mM stock (sufficient for ~500-1000 samples)

Contact us for pricing and to discuss your specific quantity requirements: +1 (714) 732-8549

Yes, DiOC2(3) is compatible with many other fluorescent dyes for multi-parameter analysis.

Common combinations:

• DiOC2(3) + Propidium Iodide (PI): Viability and membrane integrity
  • DiOC2(3)-high/PI-negative = viable cells
  • DiOC2(3)-low/PI-negative = early apoptotic
  • DiOC2(3)-low/PI-positive = late apoptotic/necrotic
• DiOC2(3) + Annexin V-PE/APC: Detailed apoptosis analysis
• DiOC2(3) + DAPI/Hoechst: Nuclear morphology and ΔΨm
• DiOC2(3) + MitoTracker Red: Dual mitochondrial labeling
• DiOC2(3) + Cell surface markers: Phenotyping with viability

Spectral compatibility:

• Green emission (497 nm) allows pairing with red/far-red dyes
• Minimal spectral overlap with PE, APC, PerCP dyes
• Compatible with 488 nm and 561/594 nm laser excitation systems

Multi-color flow cytometry:

• DiOC2(3) occupies FITC/FL1 channel
• Leaves PE, PerCP, APC channels available
• Ideal for 3-4 color panels

For specific multi-color panel design, our technical team can provide protocol recommendations.