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| Targets |
The molecular target of Copper Fluor-4 is the Cu+ (cuprous) ion. The probe is designed with a synthetic receptor that has a high selectivity and affinity for Cu+. Upon binding to Cu+, the fluorescence of the probe is quenched or turned on (typically a "turn-off" response for Cu+). The binding is reversible, allowing it to monitor dynamic changes in Cu+ concentration. This selectivity is a key feature that distinguishes it from other metal ions.
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| ln Vitro |
In vitro, the binding of Copper Fluor-4 to Cu+ results in a change in its fluorescence emission. The probe can be used to measure copper concentrations in solution. In a cuvette, addition of Cu+ to a solution of the probe will cause a decrease in fluorescence intensity. Other metal ions, such as Zn2+ or Fe2+, cause no significant change, confirming its selectivity. This property allows for the quantification of Cu+ in chemical or biological samples.
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| ln Vivo |
In vivo, Copper Fluor-4 has been shown to be useful in studying colon cancer. It can be used to image the distribution and concentration of labile copper in living cells and tissues. Its high affinity allows it to detect copper at physiological concentrations. This is important for understanding the role of copper in various diseases, including cancer, where copper levels are often dysregulated. No therapeutic effects are attributed to the probe itself.
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| Enzyme Assay |
The specificity and affinity of Copper Fluor-4 for Cu+ can be determined in a cuvette using fluorescence spectroscopy. The probe is dissolved in a suitable buffer (e.g., HEPES, pH 7.2). A fluorescence scan is performed with excitation at the optimal wavelength for the rhodol dye. A Cu+ source (e.g., [Cu(CH3CN)4]PF₆) is titrated in, and the decrease in fluorescence intensity is recorded. The Kd is calculated by fitting the binding curve. Selectivity is assessed by adding other metal ions (e.g., ZnCl2, FeSO4, CaCl2, MgCl2) at varying concentrations and measuring the change in fluorescence.
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| Cell Assay |
For cellular imaging, cells (e.g., cancer cells) are seeded on glass-bottom dishes. Copper Fluor-4 is dissolved in DMSO to a 1-10 mM stock. The culture media is replaced with media containing the probe at a final concentration of 0.5-5 uM (and often containing Pluronic F-127 to improve solubility). Cells are incubated for 30-60 minutes at 37degC, then washed. Live-cell imaging is performed on a confocal microscope to visualize the distribution of the probe (and thus, the relative concentration of Cu+). To confirm specificity, cells can be pretreated with a copper chelator (e.g., BCS) to reduce the fluorescence signal.
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| Animal Protocol |
For in vivo imaging, the probe can be injected into animal models (e.g., mice). Typically, the probe is dissolved in a water-miscible co-solvent (e.g., 5% DMSO, 30% PEG-400, and 65% saline) to achieve a concentration of 0.1-1 mg/mL. It is administered via intravenous (tail vein) or intraperitoneal injection at a dose of 0.5-2 mg/kg. After a distribution time (e.g., 30-60 minutes), the animal is imaged using an IVIS system with an appropriate filter set for the fluorophore. At the end of the study, animals are euthanized, and tissues are collected for ex vivo analysis.
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| ADME/Pharmacokinetics |
No specific pharmacokinetic data is available. Copper Fluor-4 has a molecular weight of 749.05. It is moderately lipophilic and is expected to be cell-permeable. It is soluble in DMSO and DMF. For storage, the powder should be kept at 4degC or -20degC, protected from light. Stock solutions in DMSO should be stored at -80degC for up to 6 months. Solutions in buffer may be less stable due to hydrolysis.
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| Toxicity/Toxicokinetics |
No specific toxicology data is available. Copper Fluor-4 is a research probe and has not undergone extensive toxicity testing. It should be handled as a potential health hazard. Standard laboratory safety precautions should be followed. Use a fume hood, wear gloves, lab coat, and safety goggles. Avoid inhalation and skin contact. The compound is for research use only and not for human or veterinary use.
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| References | |
| Additional Infomation |
Copper Fluor-4 (CF4) is a Cu+-specific fluorescent probe based on a rhodol scaffold. It is a research tool with extremely high affinity and selectivity for copper(I) over other metals (Kd = 2.9 × 10-¹3 M). It has been used to image copper in biological systems, including in studies of colon cancer. The compound is not a drug and has no approved therapeutic or clinical use.
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| Molecular Formula |
C38H47F3N2O2S4
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| Molecular Weight |
749.05
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| CAS # |
2365532-64-3
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| Appearance |
Solid powder
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| Density |
1.29±0.1 g/cm3(Predicted)
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| Boiling Point |
848.7±65.0 °C(Predicted)
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| LogP |
0
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| Synonyms |
CF4
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : 125 mg/mL (166.88 mM; with sonication)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3350 mL | 6.6751 mL | 13.3502 mL | |
| 5 mM | 0.2670 mL | 1.3350 mL | 2.6700 mL | |
| 10 mM | 0.1335 mL | 0.6675 mL | 1.3350 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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