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H2DCFDA [4091-99-0]

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Cat# HY-D0940-100mg

Size : 100mg

Brand : MedChemExpress

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H2DCFDA (DCFH-DA) | Redox-Sensitive Probe | MedChemExpress

H2DCFDA (DCFH-DA) is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS) (Ex/Em=488/525 nm).

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

H2DCFDA Chemical Structure

CAS No. : 4091-99-0

Customer Review

Based on 405 publication(s) in Google Scholar

394 Publications Citing Use of MCE H2DCFDA

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2022 Mar 10;e2103317. [Abstract]; CLSM images of HUVEC cells using DCFH‐DA (0.5 hours) as the fluorescent probe: Control, H2O2, H2O2+10 µg/mL1 PDA NPs, H2O2+40 µg/mL PDA NPs, H2O2+80 µg/mL PDA NPs.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Acta Pharm Sin B. 16 July 2022.; Treated HL-1 cells are washed three times and incubated with DMEM medium containing H2DCFDA (10 μM; 30 min) at 37 °C. The cells then are washed three times with warm DMEM medium and intracellular ROS production is detected by flow cytometry.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Biomaterials. 2022 Jul 21;287:121688. [Abstract]; CLSM images of 4T1 tumor cells stained with DCFH-DA (15 min) after different treatments.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Biomaterials. 2022 May;284:121513. [Abstract]; DCFH-DA (10 μM; 30 min). The ROS generation in 4T1 cells and NIH-3T3 cells incubated with various formulations for 4 h observed by CLSM.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Biomaterials. 2022 Sep 30;290:121842. [Abstract]; DCFH-DA (10 μM; 30 min) or C11-BODIPY-labeled cells are substantially increased after treatment with 1 μM FeSO4.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: J Hazard Mater. 2022 Oct 5;439:129502. [Abstract]; Representative DCFH-DA (10 μM; 30 min) staining images (Green) in PBNs exposed PSNPs and/or LPS in vivo and in vitro.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Cell. 2021 Jun 24;184(13):3528-3541.e12. [Abstract]; Stained protoplasts with H2DCFDA for the detection of ROS production and PI for PM integrity and tracked the process by live-cell imaging.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: ACS Nano. 2021 Oct 26;15(10):16286-16297. [Abstract]; CLSM image of 4T1 cells after being treated with MM, MMP, and MMTP; cells are stained with DCFH-DA (20 μM; for 1 h).

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Biomaterials. 2021 Oct;277:121103. [Abstract]; In vitro experiments are conducted to measure the cellular ROS levels in A549 cells from different treatments via flow cytometry using fluorescent probe H2DCFDA (5 μM; 30 min) to better understand the potential molecular mechanism of SOD@ARA290-HBc on radiation-induced lung injury. H2DCFDA fluorescence intensity in A549 cells significantly increased after irradiation.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Small. 2021 Aug;17(32):e2101368. [Abstract]; Laser scanning confocal fluorescence microscopy images of HT-1080 cells treated with different nanodrugs for the detection of intracellular lipid peroxidation and total ROS using C11-BODIPY and DCFH-DA (100 nM; 10 min) as the corresponding fluorescence probes.

: H2DCFDA purchased from MedChemExpress. Usage Cited in: Curr Biol. 2021 May 18;S0960-9822(21)00607-2. [Abstract]; Co-staining stigmas with H2DCFDA (50 μM; for 1-2 hours) and the cell wall indicator propidium iodide (PI) shows that ROS is located in the cytoplasm of papilla cells, near the periphery of the plasma membrane.

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Description

H2DCFDA (DCFH-DA) is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS) (Ex/Em=488/525 nm)^[1].

In Vitro

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. H2DCFDA is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use.
2. Cells are incubated with 5 μM H2DCFDA solution in PBS in the dark for 30 min at 37°C, then harvested with 0.05% trypsin-EDTA solution, suspended in a fresh medium, and immediately analyzed with flow cytometer.
3. Along with the H2DCFDA probe, if indicated, use ROS-insensitive modification of the fluorescein dye DCFDA as a positive control. The staining procedure is the same as for the H2DCFDA^[1].

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

Solvent & Solubility

In Vitro:

DMSO : 125 mg/mL (256.52 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Ethanol : 20 mg/mL (41.04 mM; Need ultrasonic)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	2.0522 mL	10.2608 mL	20.5217 mL
5 mM	0.4104 mL	2.0522 mL	4.1043 mL
10 mM	0.2052 mL	1.0261 mL	2.0522 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.

Molarity Calculator
Dilution Calculator

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

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: 2.08 mg/mL (4.27 mM); Suspended solution; Need ultrasonic

This protocol yields a suspended solution of 2.08 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 2

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.08 mg/mL (4.27 mM); Clear solution

This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Protocol 3

Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2.08 mg/mL (4.27 mM); Clear solution

This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

DMSO +

Tween-80 +

Saline

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Calculation results:

Working solution concentration: mg/mL

Method for preparing stock solution: mg drug dissolved in μL DMSO (Stock solution concentration: mg/mL).

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

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).

Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.

If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.

Purity & Documentation

Purity: 99.82%

Select Batch:

Data Sheet (277 KB) SDS (393 KB)

COA (252 KB) HNMR (228 KB) LCMS (120 KB)

Handling Instructions (2659 KB)

Dyeing Example

References

[1]. Lyublinskaya OG, et al. Redox environment in stem and differentiated cells: A quantitative approach. Redox Biol. 2017 Aug;12:758-769. [Content Brief]

Kinase Assay	ROS Measurements^[1] For the detection of intracellular ROS level ,ROS-sensitive probe H2DCFDA is used. Adherent cells (ESCs, difESCs, eMSCs, HeLa, U118) are incubated with 5 µM staining solution in PBS in the dark for 30 min at 37°C, then harvested with 0.05% trypsin-EDTA solution, suspended in a fresh medium, and immediately analyzed with flow cytometer. Lymphocytes, both control and PHA-activated, are resuspended in PBS, incubated with 5 µM of H2DCFDA in the dark for 30 min at 37°C, and immediately analyzed. Along with the H2DCFDA probe, if indicated, ROS-insensitive modification of the fluorescent dye DCFDA is used. The staining procedure is the same as for the H2DCFDA^[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.
References	[1]. Lyublinskaya OG, et al. Redox environment in stem and differentiated cells: A quantitative approach. Redox Biol. 2017 Aug;12:758-769. [Content Brief]

Kinase Assay

ROS Measurements^[1]
For the detection of intracellular ROS level ,ROS-sensitive probe H2DCFDA is used. Adherent cells (ESCs, difESCs, eMSCs, HeLa, U118) are incubated with 5 µM staining solution in PBS in the dark for 30 min at 37°C, then harvested with 0.05% trypsin-EDTA solution, suspended in a fresh medium, and immediately analyzed with flow cytometer. Lymphocytes, both control and PHA-activated, are resuspended in PBS, incubated with 5 µM of H2DCFDA in the dark for 30 min at 37°C, and immediately analyzed. Along with the H2DCFDA probe, if indicated, ROS-insensitive modification of the fluorescent dye DCFDA is used. The staining procedure is the same as for the H2DCFDA^[1].

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

References

[1]. Lyublinskaya OG, et al. Redox environment in stem and differentiated cells: A quantitative approach. Redox Biol. 2017 Aug;12:758-769. [Content Brief]

Complete Stock Solution Preparation Table

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

Ethanol / DMSO	1 mM	2.0522 mL	10.2608 mL	20.5217 mL	51.3042 mL
	5 mM	0.4104 mL	2.0522 mL	4.1043 mL	10.2608 mL
	10 mM	0.2052 mL	1.0261 mL	2.0522 mL	5.1304 mL
	15 mM	0.1368 mL	0.6841 mL	1.3681 mL	3.4203 mL
	20 mM	0.1026 mL	0.5130 mL	1.0261 mL	2.5652 mL
	25 mM	0.0821 mL	0.4104 mL	0.8209 mL	2.0522 mL
	30 mM	0.0684 mL	0.3420 mL	0.6841 mL	1.7101 mL
	40 mM	0.0513 mL	0.2565 mL	0.5130 mL	1.2826 mL
DMSO	50 mM	0.0410 mL	0.2052 mL	0.4104 mL	1.0261 mL
	60 mM	0.0342 mL	0.1710 mL	0.3420 mL	0.8551 mL
	80 mM	0.0257 mL	0.1283 mL	0.2565 mL	0.6413 mL
	100 mM	0.0205 mL	0.1026 mL	0.2052 mL	0.5130 mL

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H2DCFDA4091-99-0DCFH-DA 2',7'-Dichlorodihydrofluorescein diacetateReactive Oxygen SpeciesInhibitorinhibitorinhibit

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H2DCFDA [4091-99-0]

Cat# HY-D0940-100mg

Brand : MedChemExpress

Customer Review

H2DCFDA Related Antibodies

394 Publications Citing Use of MCE H2DCFDA

Featured Recommendations

•Related Small Molecules:

•Related Proteins:

Biological Activity

Protocol

Purity & Documentation

References

Customer Review

H2DCFDA Related Antibodies

Molarity Calculator

Dilution Calculator

Complete Stock Solution Preparation Table

H2DCFDA Related Classifications

Keywords:

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