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| Other Sizes |
| Targets |
2',7'-Dichlorofluorescein does not have a specific biological target. It functions as a chemical probe that reacts with reactive oxygen species (ROS) within cells. The compound is de-esterified intracellularly to a non-fluorescent form, which is then oxidized by ROS to a highly fluorescent product. This fluorescence can be measured to quantify ROS production, making it a valuable tool for studying oxidative stress in various biological systems.
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| ln Vitro |
In vitro, 2',7'-Dichlorofluorescein is used to measure cellular ROS production. The non-fluorescent diacetate form of the compound (DCFH-DA) is commonly used, which is cell-permeable. Once inside the cell, it is de-esterified to DCFH, which is then oxidized by ROS to the fluorescent DCF. The intensity of the fluorescence is proportional to the amount of ROS generated. It is also used for the determination of carbohydrates and hydrogen peroxide.
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| ln Vivo |
In vivo applications of 2',7'-Dichlorofluorescein are not typical, as it is primarily used in vitro for detecting ROS in cultured cells or tissue sections. It is not administered to living animals for therapeutic or diagnostic purposes. Its use is confined to laboratory research settings for studying oxidative stress and cellular redox states.
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| Enzyme Assay |
A cell-free assay for 2',7'-Dichlorofluorescein would involve measuring its fluorescence upon oxidation. In such an assay, the non-fluorescent DCFH is incubated with an oxidant, such as hydrogen peroxide, and the increase in fluorescence is measured. This can be used to detect and quantify ROS or oxidizing agents in a cell-free system. The compound's fluorescence properties, with excitation and emission spectra, can be characterized spectroscopically.
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| Cell Assay |
Cellular experiments with 2',7'-Dichlorofluorescein typically involve loading cells with the cell-permeable diacetate form (DCFH-DA). Cells are incubated with DCFH-DA, which is de-esterified intracellularly to DCFH. After washing, cells are exposed to stimuli that induce ROS production. The resulting fluorescence, indicating DCF formation, is measured using a fluorescence microplate reader, flow cytometry, or fluorescence microscopy. This assay is widely used to assess oxidative stress in various cell types.
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| Animal Protocol |
In vivo animal experiments for 2',7'-Dichlorofluorescein are not applicable, as it is a staining reagent used ex vivo on cells or tissue sections.
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| ADME/Pharmacokinetics |
2',7'-Dichlorofluorescein is a small molecule dye with a molecular weight of 401.2. Its pharmacokinetic properties are not relevant as it is not a therapeutic drug. The compound is used in vitro for research applications. It should be stored under appropriate conditions to maintain its stability and fluorescence properties.
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| Toxicity/Toxicokinetics |
Toxicity data for 2',7'-Dichlorofluorescein are not provided in the available sources. As a fluorescent probe, it is generally considered to have low toxicity for its intended research applications. However, specific toxicological information is not available. The compound is intended for research use only and not for human consumption.
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| References |
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| Additional Infomation |
2',7'-Dichlorofluorescein belongs to the 2-benzofuran class of compounds and is a fluorescent dye with functions similar to fluorescein.
2',7'-Dichlorofluorescein (CAS: 76-54-0) is an oxidation-sensitive fluorescent probe used to measure reactive oxygen species (ROS) in cells. It is the oxidized, fluorescent form of 2',7'-dichlorodihydrofluorescein (DCFH). The compound is used as an indicator and for assessing cellular antioxidant activity. It is also used for the determination of carbohydrates and hydrogen peroxide. Its molecular formula is C20H10Cl2O5. It is not a therapeutic drug and is strictly for research purposes. |
| Molecular Formula |
C20H10CL2O5
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|---|---|
| Molecular Weight |
401.1964
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| Exact Mass |
399.99
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| CAS # |
76-54-0
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| PubChem CID |
64944
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| Appearance |
Yellow to orange solid powder
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| Density |
1.8±0.1 g/cm3
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| Boiling Point |
634.4±55.0 °C at 760 mmHg
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| Melting Point |
280 °C (dec.)(lit.)
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| Flash Point |
337.4±31.5 °C
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| Vapour Pressure |
0.0±1.9 mmHg at 25°C
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| Index of Refraction |
1.797
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| LogP |
4.1
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
27
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| Complexity |
585
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
VFNKZQNIXUFLBC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H10Cl2O5/c21-13-5-11-17(7-15(13)23)26-18-8-16(24)14(22)6-12(18)20(11)10-4-2-1-3-9(10)19(25)27-20/h1-8,23-24H
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| Chemical Name |
2',7'-dichloro-3',6'-dihydroxyspiro[2-benzofuran-3,9'-xanthene]-1-one
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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 (~311.57 mM)
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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 | 2.4925 mL | 12.4626 mL | 24.9252 mL | |
| 5 mM | 0.4985 mL | 2.4925 mL | 4.9850 mL | |
| 10 mM | 0.2493 mL | 1.2463 mL | 2.4925 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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