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  4. Dihydroethidium

Dihydroethidium  (Synonyms: Hydroethidine; PD-MY 003)

Cat. No.: HY-D0079 Purity: 98.83%
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Dihydroethidium, also known as DHE, is a peroxide indicator. Dihydroethidium penetrates cell membranes to form a fluorescent protein complex with blue fluoresces. After entering the cells, Dihydroethidium is mainly localized in the cell membrane, cytoplasm and nucleus, and the staining effect is the strongest in the nucleus. Dihydroethidium produces inherent blue fluorescence with a maximum excitation wavelength of 370 nm and a maximum emission wavelength of 420 nm; after dehydrogenation, Dihydroethidium combines with RNA or DNA to produce red fluorescence with a maximum excitation wavelength of 300 nm and a maximum emission wavelength of 610 nm. 535 nm can also be used as the excitation wavelength for actual observation.

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

CAS No. : 104821-25-2

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Based on 171 publication(s) in Google Scholar

Other Forms of Dihydroethidium:

Dihydroethidium Related Antibodies

Top Publications Citing Use of Products

171 Publications Citing Use of MCE Dihydroethidium

Cell Imaging/Staining
IF

    Dihydroethidium purchased from MedChemExpress. Usage Cited in: Free Radic Biol Med. 2023 May 1:200:73-86.  [Abstract]

    The Dihydroethidium (DHE; 10 μM; 1 h), Dihydrorhodamine 123 (DHR 123, 5 μM; 1 h), FerroOrange and C11 BODIPY probes detect O2• − (red fluorescence),OONO − (green fluorescence), Fe 2+ (orange fluorescence), and lipid peroxides (red fluorescence in the unoxidized state and green fluorescence in the oxidized state), respectively, In normal lung tissue from 10 smoking patients and 10 non-smoking patients.

    Dihydroethidium purchased from MedChemExpress. Usage Cited in: Free Radic Biol Med. 2023 May 1:200:73-86.  [Abstract]

    The Dihydroethidium (DHE; 5-20 μM; 1 h）, Dihydrorhodamine 123 (DHR 123, 5 μM, 15-60 min) probes were used to detect ROS intensities in CS-induced bronchial epithelial ferroptosis. Control, and 0.5%,1%, and 2% CSE were used to stimulate BEAS-2B cells for 48 h.

    Dihydroethidium purchased from MedChemExpress. Usage Cited in: Antioxidants (Basel). 2022 Aug 19;11(8):1608.

    Superoxide anion production is measured by using the fluorescent dye Dihydroethidium (DHE). The DHE fluorescence assay detects nuclear O2●− through a chemical reaction between superoxide and DHE, whereby the latter is oxidized to ethidium.

    Dihydroethidium purchased from MedChemExpress. Usage Cited in: Oxid Med Cell Longev. 2022 Aug 4;2022:6160701.  [Abstract]

    Brain sections are incubated in 10 μM Dihydroethidium (DHE) solution for 1 h in dark at 37 °C.

    Dihydroethidium purchased from MedChemExpress. Usage Cited in: Oxid Med Cell Longev. 2021 Jun 20:2021:5546711.  [Abstract]

    HUVECs are washed with M199 and stained with Dihydroethidium (DHE, 10 μM) at 37°C with 5% CO2 for 30 min. After washing 3 times with M199, ethidium fluorescence is observed and photographed under an inversion fluorescence microscope.

    Dihydroethidium purchased from MedChemExpress. Usage Cited in: Oxid Med Cell Longev. 2021 Aug 31;2021:5608133.

    Nucleus pulposus (NP) slices are incubated with a culture medium containing 10 μM Dihydroethidium and 1 μg/mL calcein AM for one hour at 37°C. After washing with PBS, the slices are immediately viewed using an inverted confocal laser scanning microscope.

    Dihydroethidium purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2020 Apr 20;11(4):256.   [Abstract]

    The levels of ROS production in the liver tissues are determined by Dihydroethidium (DHE) staining as previously described. In brief, frozen sections of liver tissues in each group (5-μm-thick) are prepared and incubated with DHE (7.5 mM,) for 30 min in the dark at 37 °C.

    Dihydroethidium purchased from MedChemExpress. Usage Cited in: Antioxidants. 2020 Sep 18;9(9):885.  [Abstract]

    Paraffin sections (5 μm) are used to measure retinal and choroidal ROS production using a fluorescent dye with Dihydroethidium (DHE) for 20 min at 37 °C.

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    • Purity & Documentation

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    Description

    Dihydroethidium, also known as DHE, is a peroxide indicator. Dihydroethidium penetrates cell membranes to form a fluorescent protein complex with blue fluoresces. After entering the cells, Dihydroethidium is mainly localized in the cell membrane, cytoplasm and nucleus, and the staining effect is the strongest in the nucleus. Dihydroethidium produces inherent blue fluorescence with a maximum excitation wavelength of 370 nm and a maximum emission wavelength of 420 nm; after dehydrogenation, Dihydroethidium combines with RNA or DNA to produce red fluorescence with a maximum excitation wavelength of 300 nm and a maximum emission wavelength of 610 nm. 535 nm can also be used as the excitation wavelength for actual observation[1].

    In Vitro

    Guidelines (The following is our recommended protocol. This protocol only provides guidance and should be modified according to your specific needs).
    Preparation of Dihydroethidium working solution
    1.1 Preparation of stock solution
    Dissolve 1 mg Dihydroethidium in 0.317 mL DMSO to obtain a 10 mM Dihydroethidium stock solution.
    Note: It is recommended to store the stock solution at -20°C or -80°C, protected from light, and avoid repeated freeze-thaw cycles.
    1.2 Preparation of Dihydroethidium working solution
    Dilute in serum-free cell culture medium or PBS to prepare a 1-10 μM Dihydroethidium working solution.
    Note: Please adjust the concentration of the Dihydroethidium working solution according to actual conditions.
    Cell staining
    2.1 Suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and discard the supernatant. Wash twice with PBS for 5 minutes each time.
    Adherent cells: Remove the cell culture medium, add trypsin to dissociate the cells, and prepare a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes, then discard the supernatant. Wash twice with PBS for 5 minutes each time.
    2.2 Add 1 mL Dihydroethidium working solution and incubate at room temperature for 30 minutes.
    2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and discard the supernatant.
    2.4 Wash twice with PBS for 5 minutes each time.
    2.5 Resuspend the cells in serum-free cell culture medium or PBS, then detect using a fluorescence microscope (adherent cells do not require digestion) or flow cytometer.

    Storage
    -20°C, 1 year
    Protect from light

    Notes
    1. It is recommended to store the stock solution at -20°C or -80°C, protected from light, and avoid repeated freeze-thaw cycles.
    2. Please adjust the concentration of the Dihydroethidium working solution according to actual conditions.
    3. This product is for research use only.
    4. For your safety and health, please wear a lab coat and disposable gloves during operation.

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

    Dihydroethidium Related Antibodies
    Molecular Weight

    315.42

    Formula

    C21H21N3
    CAS No.
    Appearance

    Solid

    Color

    Pale purple to light pink

    Emission (Em)

    616
    Excitation (Ex)

    518
    SMILES

    NC1=CC=C2C3=C(C=C(N)C=C3)C(C4=CC=CC=C4)N(CC)C2=C1
    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 : ≥ 100 mg/mL (317.04 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    *"≥" means soluble, but saturation unknown.

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 3.1704 mL 15.8519 mL 31.7038 mL
    5 mM 0.6341 mL 3.1704 mL 6.3408 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)

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

    This equation is commonly abbreviated as: C1V1 = C2V2

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    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

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    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Calculation results:
    Working solution concentration: mg/mL
    Purity & Documentation

    Purity: 98.83%

    SDS (394 KB)

    COA (251 KB) HNMR (142 KB) RP-HPLC (163 KB) LCMS (384 KB)

    Handling Instructions (2659 KB)
    Dyeing Example
    References

    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.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    DMSO 1 mM 3.1704 mL 15.8519 mL 31.7038 mL 79.2594 mL
    5 mM 0.6341 mL 3.1704 mL 6.3408 mL 15.8519 mL
    10 mM 0.3170 mL 1.5852 mL 3.1704 mL 7.9259 mL
    15 mM 0.2114 mL 1.0568 mL 2.1136 mL 5.2840 mL
    20 mM 0.1585 mL 0.7926 mL 1.5852 mL 3.9630 mL
    25 mM 0.1268 mL 0.6341 mL 1.2682 mL 3.1704 mL
    30 mM 0.1057 mL 0.5284 mL 1.0568 mL 2.6420 mL
    40 mM 0.0793 mL 0.3963 mL 0.7926 mL 1.9815 mL
    50 mM 0.0634 mL 0.3170 mL 0.6341 mL 1.5852 mL
    60 mM 0.0528 mL 0.2642 mL 0.5284 mL 1.3210 mL
    80 mM 0.0396 mL 0.1981 mL 0.3963 mL 0.9907 mL
    100 mM 0.0317 mL 0.1585 mL 0.3170 mL 0.7926 mL
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    Dihydroethidium Related Classifications

    Help & FAQs
    • 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:

    Dihydroethidium104821-25-2Hydroethidine PD-MY 003Fluorescent DyeInhibitorinhibitorinhibit

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