| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
DCFH-DA itself is non-fluorescent, but upon entering cells, it is hydrolyzed by intracellular esterases to 2',7'-dichlorodihydrofluorescein (DCFH). DCFH is then oxidized by ROS to the highly fluorescent 2',7'-dichlorofluorescein (DCF). The fluorescence intensity is proportional to the level of ROS and NO in the cells, making it a sensitive indicator of oxidative stress.
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| ln Vitro |
In vitro, DCFH-DA is a widely used probe for detecting ROS and NO in various cell types. It is used in assays to quantify the production of reactive oxygen intermediates and to assess the degree of overall oxidative stress. The probe's fluorescence can be measured using fluorometry, flow cytometry, or fluorescence microscopy. It is a standard tool in oxidative stress research.
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| ln Vivo |
In vivo, DCFH-DA is primarily used in research applications to study oxidative stress in animal models. However, its use in vivo is limited due to its rapid metabolism and potential for non-specific oxidation. It is most commonly employed in ex vivo assays on tissue samples or in cell culture experiments.
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| Enzyme Assay |
In vitro assays for DCFH-DA typically involve incubating cells with the probe. The compound is taken up by cells, where it is deacetylated by intracellular esterases to DCFH. The cells are then stimulated with an agent that induces ROS production, and the increase in fluorescence is measured. The assay is sensitive and can detect low levels of ROS and NO.
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| Cell Assay |
Cell-based assays for DCFH-DA involve treating cells with the probe and then measuring fluorescence as a readout of oxidative stress. Cells are typically incubated with DCFH-DA for 30-60 minutes, washed, and then stimulated with a ROS-inducing agent. The fluorescence is measured using a fluorometer, flow cytometer, or fluorescence microscope. The assay is widely used to study the effects of various compounds on cellular oxidative stress.
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| Animal Protocol |
In vivo animal studies for DCFH-DA are limited, but the probe can be used in ex vivo assays on tissue samples from treated animals. For example, tissue homogenates can be incubated with DCFH-DA, and fluorescence can be measured to assess ROS levels. This approach is used to study the effects of drugs or disease on oxidative stress in various organs.
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| ADME/Pharmacokinetics |
Diacetyldichlorofluorescein has a molecular weight of 485.26 g/mol and a molecular formula of C24H14Cl2O7. It is soluble in DMSO at approximately 33.33 mg/mL (68.68 mM). The compound is light-sensitive and should be protected from light during transportation and storage. For storage, the powder should be kept at -20°C for up to 3 years or at 4°C for up to 2 years; in solution, it can be stored at -80°C for 6 months or at -20°C for 1 month.
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| Toxicity/Toxicokinetics |
The toxicological profile of DCFH-DA is consistent with that of other fluorescent probes. It is generally considered non-toxic at the concentrations used in assays. However, as with any chemical reagent, appropriate safety precautions should be taken when handling the compound. Its use is limited to research applications.
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| References | |
| Additional Infomation |
Diacetyldichlorofluorescein is a research compound with no approved human therapeutic use. It is a widely used fluorescent probe for the detection of ROS and NO in cells and tissues. It is a valuable tool for studying oxidative stress, inflammation, and the effects of various compounds on cellular redox status. The probe is used in a wide range of research fields, including cell biology, pharmacology, and toxicology.
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| Molecular Formula |
C24H14CL2O7
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|---|---|
| Molecular Weight |
485.26
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| Exact Mass |
484.012
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| CAS # |
2044-85-1
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| PubChem CID |
104913
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| Appearance |
Light yellow to orange solid powder
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| Density |
1.59 g/cm3
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| Boiling Point |
657.9ºC at 760 mmHg
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| Melting Point |
232-234ºC(lit.)
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| Flash Point |
239.2ºC
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| Vapour Pressure |
3.49E-17mmHg at 25°C
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| Index of Refraction |
1.692
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| LogP |
5.412
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
33
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| Complexity |
775
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
VQVUBYASAICPFU-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H14Cl2O7/c1-11(27)30-21-9-19-15(7-17(21)25)24(14-6-4-3-5-13(14)23(29)33-24)16-8-18(26)22(31-12(2)28)10-20(16)32-19/h3-10H,1-2H3
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| Chemical Name |
(6'-acetyloxy-2',7'-dichloro-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl) acetate
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| Synonyms |
Leuco-dcf diacetate Dcfh-DA Diacetyldichlorofluorescein
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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 : ~33.33 mg/mL (~68.68 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.15 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0608 mL | 10.3038 mL | 20.6075 mL | |
| 5 mM | 0.4122 mL | 2.0608 mL | 4.1215 mL | |
| 10 mM | 0.2061 mL | 1.0304 mL | 2.0608 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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