| Size | Price | Stock | Qty |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g | |||
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| Targets |
The target of 2',7'-Dichlorodihydrofluorescein is not a biological receptor but rather reactive oxygen species (ROS) themselves. The probe is oxidized by various ROS including hydroxyl radicals (·OH), hydrogen peroxide (H₂O₂), peroxynitrite (ONOO⁻), and other reactive species. The oxidation reaction converts the non-fluorescent reduced form (DCFH) to the highly fluorescent oxidized form (DCF). This property makes it a valuable tool for measuring intracellular oxidative stress levels.
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| ln Vitro |
In vitro, 2',7'-Dichlorodihydrofluorescein is used as a fluorescent probe to detect ROS in cell-free systems and cell cultures. The non-fluorescent probe is oxidized by ROS to produce the highly fluorescent DCF. The fluorescence intensity is measured and correlates with ROS concentration. The probe has an excitation wavelength of 485-500 nm and an emission wavelength of 515-530 nm. It is widely used in oxidative stress and cell damage research.
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| ln Vivo |
Specific in vivo data for 2',7'-Dichlorodihydrofluorescein are limited as it is primarily used as an in vitro and ex vivo probe. However, the compound can be used to measure ROS levels in tissues ex vivo after administration to animals. It is also used in live-cell imaging and flow cytometry applications. The probe's ability to detect ROS in biological systems makes it valuable for studying oxidative stress in disease models, though its use is typically limited to in vitro or ex vivo applications.
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| Enzyme Assay |
The ROS detection assay is performed by incubating 2',7'-Dichlorodihydrofluorescein with samples containing ROS. The non-fluorescent DCFH is oxidized to fluorescent DCF. Fluorescence is measured at excitation 485-500 nm and emission 515-530 nm. In cell-free systems, known concentrations of H₂O₂ or other ROS are used to generate a standard curve. The probe is typically dissolved in DMSO or methanol and diluted in buffer before use. The reaction is typically carried out at 37°C for 30-60 minutes.
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| Cell Assay |
For cellular studies, cells are cultured in appropriate media and loaded with 2',7'-Dichlorodihydrofluorescein. The non-fluorescent probe (typically 10-50 μM) is added to the culture medium and incubated at 37°C for 30-60 minutes to allow cellular uptake. After washing to remove extracellular probe, cells are treated with test compounds or exposed to oxidative stress inducers. Fluorescence is measured using a fluorescence microplate reader, flow cytometry, or fluorescence microscopy. The increase in fluorescence indicates ROS production.
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| Animal Protocol |
In vivo studies using 2',7'-Dichlorodihydrofluorescein are typically performed ex vivo. Animals are treated with test compounds or exposed to oxidative stress conditions, and tissues are harvested. Tissue sections or homogenates are incubated with the probe, and fluorescence is measured to assess ROS levels. Alternatively, the probe can be administered directly to animals for in vivo imaging applications, though this is less common due to rapid metabolism and clearance of the probe.
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| ADME/Pharmacokinetics |
Specific pharmacokinetic data for 2',7'-Dichlorodihydrofluorescein are not typically reported as it is a probe rather than a therapeutic compound. The compound has a molecular weight of 403.21 g/mol and is a small molecule that can penetrate cell membranes. It is typically used at concentrations of 1-50 μM in cell culture experiments. The probe should be protected from light and stored at -20°C to prevent oxidation.
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| Toxicity/Toxicokinetics |
2',7'-Dichlorodihydrofluorescein is considered non-toxic at the concentrations typically used for ROS detection (1-50 μM). However, as with any chemical probe, appropriate safety precautions should be taken. The compound should be handled with gloves and eye protection. It should be protected from light to prevent premature oxidation. No significant toxicity has been reported at standard working concentrations. The compound is for research use only and not for human consumption.
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| Additional Infomation |
See also: 2',7'-Dichlorofluorescein (note moved to).
2',7'-Dichlorodihydrofluorescein (DCFH) is a non-fluorescent ROS probe that is oxidized by intracellular ROS to produce highly fluorescent DCF. It is widely used for quantitative detection of oxidative stress in cells and biological systems. Excitation/emission: 485-500/515-530 nm. No clinical applications exist. For research use only. |
| Molecular Formula |
C20H12O5CL2
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|---|---|
| Molecular Weight |
403.21228
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| Exact Mass |
402.006
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| CAS # |
106070-31-9
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| PubChem CID |
14598431
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| Appearance |
Pink to orange solid powder
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| Density |
1.583g/cm3
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| Boiling Point |
553.7ºC at 760mmHg
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| Flash Point |
288.7ºC
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| Vapour Pressure |
4.26E-13mmHg at 25°C
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| Index of Refraction |
1.717
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| LogP |
5.388
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
27
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| Complexity |
531
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ClC1C(O)=CC2=C(C(C3C=CC=CC=3C(O)=O)C3C(O2)=CC(O)=C(Cl)C=3)C=1
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| InChi Key |
XDFNWJDGWJVGGN-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H12Cl2O5/c21-13-5-11-17(7-15(13)23)27-18-8-16(24)14(22)6-12(18)19(11)9-3-1-2-4-10(9)20(25)26/h1-8,19,23-24H,(H,25,26)
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| Chemical Name |
2-(2,7-dichloro-3,6-dihydroxy-9H-xanthen-9-yl)benzoic acid
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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 |
| 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 : ~250 mg/mL (~620.02 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 20.83 mg/mL (51.66 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 208.3 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (5.16 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (5.16 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4801 mL | 12.4005 mL | 24.8010 mL | |
| 5 mM | 0.4960 mL | 2.4801 mL | 4.9602 mL | |
| 10 mM | 0.2480 mL | 1.2400 mL | 2.4801 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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