Diacetyldichlorofluorescein (DCFH-DA; 2′,7′-Dichlorofluorescein diacetate) is a cell-permeable fluorescent probe and stable storage form of dichlorofluorescein. It can be used to detect the generation of reactive oxygen intermediates and for assessing the overall oxidative stress in toxicological phenomenon.

Physicochemical Properties

Molecular Formula	C24H14CL2O7
Molecular Weight	485.26
Exact Mass	484.012
CAS #	2044-85-1
PubChem CID	104913
Appearance	Light yellow to orange solid powder
Density	1.59 g/cm3
Boiling Point	657.9ºC at 760 mmHg
Melting Point	232-234ºC(lit.)
Flash Point	239.2ºC
Vapour Pressure	3.49E-17mmHg at 25°C
Index of Refraction	1.692
LogP	5.412
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	4
Heavy Atom Count	33
Complexity	775
Defined Atom Stereocenter Count	0
InChi Key	VQVUBYASAICPFU-UHFFFAOYSA-N
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
Chemical Name	(6'-acetyloxy-2',7'-dichloro-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl) acetate
Synonyms	Leuco-dcf diacetate Dcfh-DA Diacetyldichlorofluorescein
HS Tariff Code	2934.99.9001
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)

Biological Activity

References

[1]. Active oxygen chemistry within the liposomal bilayer. Part IV: Locating 2',7'-dichlorofluorescein (DCF), 2',7'-dichlorodihydrofluorescein (DCFH) and 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) in the lipid bilayer. Chem Phys Lipids. 2004 Aug;131(1):123-33.

Solubility Data

Solubility (In Vitro)

DMSO : ~33.33 mg/mL (~68.68 mM)

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.