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Hoechst 33342  (Synonyms: bisBenzimide H 33342; HOE 33342)

Cat. No.: HY-15559 Purity: 99.94%
COA Handling Instructions

Hoechst 33342 is a marker dye in Hoechst series. Hoechst is A live nuclear marker dye. Hoechst binds to the grooves in the DNA double strand, which tends to be A/ T-rich DNA strand. Although it binds to all nucleic acids, the A/ T-rich double strand DNA significantly enhances fluorescence intensity Therefore,Hoechst dye can be used for living cell labeling. The fluorescence intensity of Hoechst dye increases with the increase of pH of solution.

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

Hoechst 33342 Chemical Structure

Hoechst 33342 Chemical Structure

CAS No. : 23491-52-3

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Customer Review

Based on 59 publication(s) in Google Scholar

Other Forms of Hoechst 33342:

Top Publications Citing Use of Products

49 Publications Citing Use of MCE Hoechst 33342

IF

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: Bioact Mater. 2022 Mar 17;18:91-103.  [Abstract]

    The fluorescent images of HUVECs stained with NO fluorescent probe (DAF-FM DA) and Hoechst 33342 after being treated with UCATS under dark conditions or 808 nm NIR irradiation.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: Chem Eng J. 15 December 2022, 138004.

    Apoptotic cells in four groups were stained with Hoechst 33342 (for 10 min) reflected by bright blue nuclei, including hP-MSCs cultured on noncoated, L-Gel-coated (10 nM), and D-Gel coated (10 nM) plates under exposure of 500 μM H2O2 for 5 h except for the normal group, and PBS as control.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: J Lipid Res. 2022 May;63(5):100207.  [Abstract]

    complete medium containing 1 μg/mL BODIPY 500/510 for 3 h and then 2 μg/mL Hoechst 33342 for an additional 2 min.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: Theranostics. 2021 Mar 4;11(10):4655-4671.  [Abstract]

    Hoechst 33342 (10 µg/mL) is added to the medium of cultured megakaryocytes for 2 h at 37°C. Cells are stained with the directly coupled monoclonal antibodies PE-anti-CD41 and APC-anti-Mpl for 30 min at 4°C. Ploidy is measured in the CD41+Mpl+ cell population.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: J Cell Biol. 2021 Oct 4;220(10):e202011153.  [Abstract]

    Chromatin is briefly counterstained with Hoechst 33342 (5 µg/mL) before imaging.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: Aging Cell. 2021 Jul;20(7):e13382.  [Abstract]

    BM cells are incubated with 5 μg/mL Hoechst 33342 in Hank balanced salt solution containing 20 mM HEPES, 5 mM glucose, and 10% fetal bovine serum at 37°C for 45 min, followed by an additional 45 min of incubation with 1 μg/ml Pyronin Y.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: Stem Cell Res Ther. 2021 Jan 22;12(1):77.  [Abstract]

    To evaluate the anti-apoptosis property of extracellular vesicles from hP-MSCs, the Hoechst 33342 (for 10 min) is used to distinguish the apoptotic cells in the three different groups.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: Acta Biomater. 2020 Sep 1;113:289-304.  [Abstract]

    Hoechst 33342 staining (for 6 h) is used to distinguish apoptotic changes in H2O2-treated hP-MSCs.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: ACS Appl Mater Interfaces. 2018 Sep 12;10(36):30081-30091.  [Abstract]

    Hoechst 33342 staining shows the survival of HUVECs administrated 100 μg/mL exosomes or CS-Exo upon exposure to 400 μM H2O2.

    Hoechst 33342 purchased from MedChemExpress. Usage Cited in: Chinese Pharmacological Bulletin. 2018 May; 34(5): 620-626.

    PC12 cells are treated with 1.5 mM Methylglyoxal (MG) for 24 h before pretreatment with Butein (2.5, 5 μM) for 1 h. Propidium iodide (PI) and Hoechst 33342 are used to determine cell apoptosis.
    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

    • Customer Review

    Description

    Hoechst 33342 is a marker dye in Hoechst series. Hoechst is A live nuclear marker dye. Hoechst binds to the grooves in the DNA double strand, which tends to be A/ T-rich DNA strand. Although it binds to all nucleic acids, the A/ T-rich double strand DNA significantly enhances fluorescence intensity Therefore,Hoechst dye can be used for living cell labeling. The fluorescence intensity of Hoechst dye increases with the increase of pH of solution[1].

    IC50 & Target

    Dye reagent[1]
    DNA Stain[1]

    In Vitro

    General Protocol
    Preparation of Hoechst working solution
    1.1 Preparation of the stock solution
    Dissolve 10 mg of in 5 mL DMSO
    Note: It is recommended to store the stock solution at 4℃ or -20℃ away from light and avoid repetitive freeze-thaw cycles.
    1.2 Preparation of Hoechst working solution
    Dilute the stock solution in serum-free cell culture medium or PBS to obtain final concentration 10 μg/mL Hoechst working solution.
    Note: Please adjust the concentration of Hoechst working solution according to the actual situation.
    1.Cell staining
    2.1 Suspension cells(6-well plate)
    a. Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time. The cell density is 1×106/mL.
    b. Add 1 mL of working solution, and then incubate at room temperature for 3-10 minutes.
    c. Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
    d. Wash twice with PBS, 5 minutes each time.
    e. Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
    2.2 Adherent cells
    a. Culture adherent cells on sterile coverslips.
    b. Remove the coverslip from the medium and aspirate excess medium.
    c. Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 3-10 minutes.
    d. Wash twice with medium, 5 minutes each time.Observation by fluorescence microscopy or flow cytometry.

    Precautions
    1. Please adjust the concentration of Hoechst working solution according to the actual situation.
    2. This product is for R&D use only, not for drug, household, or other uses.
    3. 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.

    Molecular Weight

    452.55

    Formula

    C27H28N6O

    CAS No.
    Appearance

    Solid

    Color

    Brown to breen

    Emission (Em)

    452

    Excitation (Ex)

    356

    SMILES

    CN1CCN(C2=CC=C3C(N=C(C4=CC=C5C(N=C(C6=CC=C(OCC)C=C6)N5)=C4)N3)=C2)CC1

    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 : 6.25 mg/mL (13.81 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    H2O : < 0.1 mg/mL (insoluble)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 2.2097 mL 11.0485 mL 22.0970 mL
    5 mM 0.4419 mL 2.2097 mL 4.4194 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: ≥ 0.5 mg/mL (1.10 mM); Clear solution

      This protocol yields a clear solution of ≥ 0.5 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (5.0 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: ≥ 0.5 mg/mL (1.10 mM); Clear solution

      This protocol yields a clear solution of ≥ 0.5 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (5.0 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

    Purity: 99.94%

    Dyeing Example
    References
    Cell Assay
    [1]

    Labeling Nuclear DNA with Hoechst 33342[1] Step 1, Dilute the Hoechst stock solution 1:100 in H2O for use in labeling. Step 2, Aspirate the cell medium from cells grown on coverslips. Rinse the cells three times with PBS+. Step 3, Incubate the cells in the Hoechst labeling solution (from Step 1) for 10-30 min at room temperature. Step 4, Aspirate the labeling solution. Rinse the cells three times in PBS+. Step 5, Mount the coverslips. Step 6, Image the cells (λex ~353 nm, λem ~483 nm for Hoechst 33342)[1].

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

    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 2.2097 mL 11.0485 mL 22.0970 mL 55.2425 mL
    5 mM 0.4419 mL 2.2097 mL 4.4194 mL 11.0485 mL
    10 mM 0.2210 mL 1.1049 mL 2.2097 mL 5.5243 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.

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