2′,7′-Dichlorofluorescein diacetate (Synonyms: DCFH2-DA)

Name: 2′,7′-Dichlorofluorescein diacetate
Brand: MedChemExpress
SKU: HY-126793
Rating: 5.0 (23 reviews)

Cat. No.: HY-126793 Purity: 99.16%: Data Sheet
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2′,7′-Dichlorofluorescein diacetate (DCFH2-DA) is a cell-permeable fluorescent probe. 2′,7′-Dichlorofluorescein diacetate can be used to detect the generation of reactive oxygen intermediates and for assessing the overall oxidative stress in toxicological phenomenon.

For research use only. We do not sell to patients.

CAS No. : 2044-85-1

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10 mM * 1 mL in DMSO ready for reconstitution	In-stock	Estimated Time of Arrival: December 31
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10 mM * 1 mL in DMSO	In-stock	Estimated Time of Arrival: December 31
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Based on 23 publication(s) in Google Scholar

Other Forms of 2′,7′-Dichlorofluorescein diacetate:

2′,7′-Dichlorofluorescein diacetate (solution) Get quote

23 Publications Citing Use of MCE 2′,7′-Dichlorofluorescein diacetate

Cell Imaging/Staining

: 2′,7′-Dichlorofluorescein diacetate purchased from MedChemExpress. Usage Cited in: Redox Biol. 2024 Dec 25:79:103482. [Abstract]; ROS levels were detected with the 2′,7′-Dichlorofluorescein diacetate (DCFH-DA, 10 μM) fluorescent probe in HaCaT cells.

: 2′,7′-Dichlorofluorescein diacetate purchased from MedChemExpress. Usage Cited in: Food Chem. 2025 Jun 9:490:145079. [Abstract]; The impact of RPPH-1 treatment on the activities of reactive oxygen species (ROS) in C. elegans. In this experiment, 2′,7′-dichlorofluorescein diacetate (DCFH2-DA, 10 mM; 1 h) was used to detect ROS.

: 2′,7′-Dichlorofluorescein diacetate purchased from MedChemExpress. Usage Cited in: Phytomedicine. 2025 Oct:146:157098. [Abstract]; ROS levels in A549 cells after 4 h of Flp-A treatment were measured using a ROS detection probe (2′,7′-dichlorofluorescein diacetate (DCFH2-DA)) (scale bar = 100 μm).

: 2′,7′-Dichlorofluorescein diacetate purchased from MedChemExpress. Usage Cited in: Adv Mater. 2024 Nov;36(45):e2409618. [Abstract]; Intracellular ROS level of Hepa 1-6 cells after different treatments. This experiment utilized the 2′,7′-dichlorofluorescein diacetate (DCFH-DA) (10 μM; 25 min) probe to visualize and characterize the generation process of ROS.

: 2′,7′-Dichlorofluorescein diacetate purchased from MedChemExpress. Usage Cited in: Food Hydrocoll. 93 (2019) 261-269.; Images of control group (Control), H₂O₂ treated group, H₂O₂ treated and then digestion fluid of zein particle encapsulated resveratrol treated group (ZP), digestion fluid of free resveratrol group treated group (Free Res), and digestion fluid of physical mixture treated group (PM). The higher the fluorescence intensity, the higher the ROS level. The digestion fluid samples had been subjected to 120 min stimulated gastric digestion and 240 min small intestinal digestion prior to analysis.
H₂O₂ was used to cause oxidative damage to the cells, and 2′,7′-Dichlorofluorescein diacetate (DCFH-DA) was used as a fluorescence probe to monitor the intracellular ROS level. A very weak fluorescence signal was observed for the control, which indicated that there was little ROS in the HepG2 cells themselves.

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Description

In Vitro

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
Using 2',7'-Dichlorofluorescein as a fluorescent probe to quantify intracellular ROS^[2]
Experimental Reagents
20,70-dichlorofluorescein diacetate (DCFH2-DA): 100 nM.
CoCl₂: 20 mM, used as an extracellular DCF fluorescence quencher.
Cell lines:
K562: Human erythromyelogenous leukemia cell line (drug-sensitive).
K562/adr: Drug-resistant subline of K562.
GLC4: Human small cell lung carcinoma cell line (drug-sensitive).
GLC4/adr: Drug-resistant subline of GLC4.
Culture medium: RPMI 1640, supplemented with 10% fetal bovine serum (FBS).
Experimental Procedure
1. Cell Culture:
(1) Seed K562, K562/adr, GLC4, and GLC4/adr cells into RPMI 1640 medium supplemented with 10% FBS, and culture in a 37°C, 5% CO2 incubator.
(2) The cell density should be 5×10⁵ cells/mL, and proceed to the next step after 24 hours.
2. ROS Measurement:
(1) Treat the cells: Add 20 μM CoCl₂ (extracellular fluorescence quencher) and 100 nM DCHF-DA, and continue incubation for 30 minutes to ensure sufficient oxidation of DCHF by ROS.
(2) Measure fluorescence: Use a Perkin-Elmer LS-50B fluorimeter, set the excitation wavelength to 502 nm, and the emission wavelength to 523 nm, to monitor the fluorescence intensity of DCF.
(3) Data analysis:
Fit the kinetic model based on fluorescence intensity changes to calculate the deacetylation rate constant (ke) of DCHF-DA and the rate constant of ROS oxidation (k2[ROSi]). Use this model to quantitatively analyze ROS concentrations in different cell lines.
Experimental Results
1. ROS concentration: ROS concentrations vary between different cell lines. The average ROS concentration in K562 cells is 0.67 nM, 0.60 nM in K562/adr cells, 0.66 nM in GLC4 cells, and 0.81 nM in GLC4/adr cells.
2. Experimental validation: The use of CoCl₂ as a quencher effectively removes external ROS signals, ensuring that the obtained DCF fluorescence signal accurately reflects intracellular ROS concentration.
Note: Drug-resistant cell lines (such as K562/adr and GLC4/adr) show higher ROS levels, suggesting that ROS accumulation may be involved in the mechanisms of drug resistance.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

485.27

Formula

C₂₄H₁₄Cl₂O₇

CAS No.

2044-85-1

Appearance

Solid

Color

Off-white to light yellow

Emission (Em)

525

Excitation (Ex)

496

SMILES

O=C1OC2(C3=C(OC4=C2C=C(Cl)C(OC(C)=O)=C4)C=C(OC(C)=O)C(Cl)=C3)C5=C1C=CC=C5

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility

In Vitro:

DMSO : 33.33 mg/mL (68.68 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	2.0607 mL	10.3035 mL	20.6071 mL
5 mM	0.4121 mL	2.0607 mL	4.1214 mL
10 mM	0.2061 mL	1.0304 mL	2.0607 mL

View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

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Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

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Purity & Documentation

Purity: 99.16%

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Data Sheet (280 KB) SDS (393 KB)

COA (255 KB) HNMR (265 KB) RP-HPLC (260 KB) LCMS (103 KB)

Handling Instructions (2659 KB)

References

[1]. Afri M, et al. 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. [Content Brief]

[2]. Loetchutinat C, et al. Spectrofluorometric determination of intracellular levels of reactive oxygen species in drug-sensitive and drug-resistant cancer cells using the 2′, 7′-dichlorofluorescein diacetate assay[J]. Radiation Physics and Chemistry, 2005, 72(2-3): 323-331.

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

DMSO	1 mM	2.0607 mL	10.3035 mL	20.6071 mL	51.5177 mL
	5 mM	0.4121 mL	2.0607 mL	4.1214 mL	10.3035 mL
	10 mM	0.2061 mL	1.0304 mL	2.0607 mL	5.1518 mL
	15 mM	0.1374 mL	0.6869 mL	1.3738 mL	3.4345 mL
	20 mM	0.1030 mL	0.5152 mL	1.0304 mL	2.5759 mL
	25 mM	0.0824 mL	0.4121 mL	0.8243 mL	2.0607 mL
	30 mM	0.0687 mL	0.3435 mL	0.6869 mL	1.7173 mL
	40 mM	0.0515 mL	0.2576 mL	0.5152 mL	1.2879 mL
	50 mM	0.0412 mL	0.2061 mL	0.4121 mL	1.0304 mL
	60 mM	0.0343 mL	0.1717 mL	0.3435 mL	0.8586 mL

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Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

2′,7′-Dichlorofluorescein diacetate2044-85-1DCFH2-DAFluorescent DyeReactive Oxygen Species (ROS)cell-permeablereactive oxygenROSoxidative stressInhibitorinhibitorinhibit

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2′,7′-Dichlorofluorescein diacetate (Synonyms: DCFH2-DA)

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Other Forms of 2′,7′-Dichlorofluorescein diacetate:

2′,7′-Dichlorofluorescein diacetate Related Antibodies

23 Publications Citing Use of MCE 2′,7′-Dichlorofluorescein diacetate

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2′,7′-Dichlorofluorescein diacetate Related Antibodies

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Complete Stock Solution Preparation Table

2′,7′-Dichlorofluorescein diacetate Related Classifications

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