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  4. Dihydroethidium

Dihydroethidium  (Synonyms: Hydroethidine; PD-MY 003)

Cat. No.: HY-D0079 Purity: 98.71%
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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.

Dihydroethidium Chemical Structure

Dihydroethidium Chemical Structure

CAS No. : 104821-25-2

Size Price Stock Quantity
5 mg USD 40 In-stock
10 mg USD 66 In-stock
25 mg USD 145 In-stock
50 mg USD 232 In-stock
100 mg USD 371 In-stock
200 mg   Get quote  
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Based on 81 publication(s) in Google Scholar

Dihydroethidium Related Antibodies

Top Publications Citing Use of Products

74 Publications Citing Use of MCE Dihydroethidium

IF

    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.

    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 Mar 20;2021:6699054.

    H9c2 myoblasts are treated with IL-6 and Raloxifene as described above and then are incubated with 10 μM Dihydroethidium (DHE) at 37°C for 30 minutes in the dark and washed twice with PBS.

    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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    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

    General Protocol
    Preparation of Dihydroethidium working solution
    1.1 Preparation of the stock solution
    Dissolve 1 mg of Dihydroethidium in 0.31 mL of DMSO to obtain 10 mM of Dihydroethidium.
    Note: It is recommended to store the stock solution at -20°C or -80°C away from light and avoid repetitive freeze-thaw cycles.
    1.2 Preparation of Dihydroethidium working solution
    Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of Dihydroethidium working solution.
    Note: Please adjust the concentration of Dihydroethidium working solution according to the actual situation.
    Cell staining
    2.1 For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
    For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
    2.2 Add 1 mL of Dihydroethidium working solution, and then incubate at room temperature for 30 minutes.
    2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
    2.4 Wash twice with PBS, 5 minutes each time.
    2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

    Storage
    -20°C, 1 year
    Protect from light

    Precautions
    1. It is recommended to store the stock solution at -20℃ or -80℃ away from light and avoid repetitive freeze-thaw cycles.
    2. Please adjust the concentration of Dihydroethidium working solution according to the actual situation.
    3. This product is for R&D use only, not for drug, household, or other uses.
    4. For your safety and health, please wear a lab coat and disposable gloves to operate.

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

    Dihydroethidium Related Antibodies
    Molecular Weight

    315.41

    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 (317.05 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 3.1705 mL 15.8524 mL 31.7048 mL
    5 mM 0.6341 mL 3.1705 mL 6.3410 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)

    Mass
    =
    Concentration
    ×
    Volume
    ×
    Molecular Weight *

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

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

    ×
    Volume (start)

    V1

    =
    Concentration (final)

    C2

    ×
    Volume (final)

    V2

    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: 2.5 mg/mL (7.93 mM); Suspended solution; Need ultrasonic

      This protocol yields a suspended solution of 2.5 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 (25.0 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: 2.5 mg/mL (7.93 mM); Suspended solution; Need ultrasonic

      This protocol yields a suspended solution of 2.5 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 (25.0 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.
    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

    Animal weight
    (per animal)

    g

    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
    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 3.1705 mL 15.8524 mL 31.7048 mL 79.2619 mL
    5 mM 0.6341 mL 3.1705 mL 6.3410 mL 15.8524 mL
    10 mM 0.3170 mL 1.5852 mL 3.1705 mL 7.9262 mL
    15 mM 0.2114 mL 1.0568 mL 2.1137 mL 5.2841 mL
    20 mM 0.1585 mL 0.7926 mL 1.5852 mL 3.9631 mL
    25 mM 0.1268 mL 0.6341 mL 1.2682 mL 3.1705 mL
    30 mM 0.1057 mL 0.5284 mL 1.0568 mL 2.6421 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 0.1982 mL 0.3963 mL 0.9908 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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