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> DAPI (dihydrochloride) [28718-90-3]

DAPI (dihydrochloride) [28718-90-3]

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DAPI dihydrochloride is a DAPI dye. DAPI is a fluorescent dye that binds strongly to DNA. It binds to the AT base pair of the double-stranded DNA minor groove, and one DAPI molecule can occupy three base pair positions. The fluorescence intensity of DAPI molecules bound to double-stranded DNA is increased by about 20 times, and it is commonly observed with fluorescence microscopy, and the amount of DNA can be determined based on the intensity of fluorescence. In addition, because DAPI can pass through intact cell membranes, it can be used to stain both live and fixed cells.

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DAPI dihydrochloride Estructura química

DAPI dihydrochloride Estructura química

No. CAS : 28718-90-3

This product is a controlled substance and not for sale in your territory.

Based on 57 publication(s) in Google Scholar

Other Forms of DAPI dihydrochloride:

  • DAPI dilactate Obtener un presupuesto

    DAPI dihydrochloride purchased from MedChemExpress. Usage Cited in: Pharmaceuticals. 2022 Jun 6;15(6):721.

    The nuclei are counterstained with DAPI (10 μM) in the dark for 10 min at room temperature. Fluorescence is monitored using confocal microscope, and foci of phospho-H2A.X was quantified using ImageJ software.

    DAPI dihydrochloride purchased from MedChemExpress. Usage Cited in: Acta Pharmacol Sin. 2021 Jul;42(7):1111-1123.  [Abstract]

    Mice intestine tissues sections were incubated with a TUNEL reaction mixture for 1 h at 37 °C in the dark. Next, the sections are stained with 0.1 μg/mL DAPI.

    DAPI dihydrochloride purchased from MedChemExpress. Usage Cited in: Oncotargets Ther. 2020 Oct 6;13:9941-9952.  [Abstract]

    Human GC MGC-803 and NCI-N87 cells are seeded into 24-well plates with glass slide for 24h in RPMI-1640 medium. And 0.1% DAPI staining is performed against light for 30 minutes.

    DAPI dihydrochloride purchased from MedChemExpress. Usage Cited in: Acta Biomater. 2019 Sep 15;96:468-479.  [Abstract]

    CLSM observations of tested Cy5-nanocomplexes incubated with RAW 264.7 macrophages show the fluorescent distributions of DAPI, nanocomplexes, and ROS (DCFH-DA) in the presence and absence NIR treatment.

    DAPI dihydrochloride purchased from MedChemExpress. Usage Cited in: Phytomedicine. 2020 Mar;68:153146.  [Abstract]

    MC3T3-E1 cells incubated with 1 μg/ml DAPI dihydrochloride for 5 min and observed them under a fluorescence microscope. Effects of sweroside on ER-α and GPR30 expressions by immunofluorescence, white and yellow arrows mean target protein distribution in the cell nuclei and membrane, respectively.
    Descripciòn

    DAPI dihydrochloride is a DAPI dye. DAPI is a fluorescent dye that binds strongly to DNA. It binds to the AT base pair of the double-stranded DNA minor groove, and one DAPI molecule can occupy three base pair positions. The fluorescence intensity of DAPI molecules bound to double-stranded DNA is increased by about 20 times, and it is commonly observed with fluorescence microscopy, and the amount of DNA can be determined based on the intensity of fluorescence. In addition, because DAPI can pass through intact cell membranes, it can be used to stain both live and fixed cells[1].

    In Vitro

    General Protocol
    1. Preparation of DAPI working solution
    1.1 Preparation of the stock solution
    Dissolve 1 mg DAPI in 1 mL ddH2O to obtain 1 mg/mL of stock solution.
    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 DAPI working solution
    Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 ug/mL of working solution.
    Note: Please adjust the concentration of DAPI working solution according to the actual situation.
    2. Cell staining
    2.1 Suspension cells（6-well plate）
    a. Centrifuge at 1000 g at 4°C 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°C 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.

    Storage
    -20°C, 1 year. Protect from light

    Precautions 1. Please adjust the concentration of DAPI 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.

    DAPI dihydrochloride Related Antibodies
    Peso molecular

    350.25

    Fòrmula

    C16H17Cl2N5
    No. CAS

    28718-90-3
    Appearance

    Solid

    Color

    Light yellow to green yellow

    Emission (Em)

    451
    Excitation (Ex)

    356
    SMILES

    N=C(C1=CC2=C(C=C1)C=C(C3=CC=C(C(N)=N)C=C3)N2)N.01Cl.01Cl
    Envío

    Room temperature in continental US; may vary elsewhere.
    Almacenamiento

    -20°C, sealed storage, away from moisture and light

    *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

    Solvente y solubilidad
    In Vitro: 

    DMSO : 75 mg/mL (214.13 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    H2O : 5.56 mg/mL (15.87 mM; ultrasonic and warming and heat to 60°C)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 2.8551 mL 14.2755 mL 28.5510 mL
    5 mM 0.5710 mL 2.8551 mL 5.7102 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 (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    * Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

    • Calculadora de molaridad

    • Calculadora de dilución

    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: ≥ 2.08 mg/mL (5.94 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 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 (5.94 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.
    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 (sealed storage, away from moisture and 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.
    Pureza y Documentación

    Purity: 99.30%

    COA (246 KB) LCMS (94 KB)

    Instrucciones de manejo (2659 KB)
    Dyeing Example
    • Description: DAPI dihydrochloride is a DNA-specific probe with blue fluorescence.
      Method: For cell staining.
      1. Cells are cultured in Dulbecco’s modified Eagle medium (DMEM), and supplemented with 10% fetal bovine serum.
      2. Cells are expanded in an incubator for 12 h at 37℃ with 5% CO2 and then seeded onto confocal dishes.
      3. Cells are grown until confluence and washed with Hank’s balanced salt solution (HBSS).
      4. Cells are stained with DAPI dihydrochloride and imaged by a confocal laser scanning microscopy.
    • Description: DAPI dihydrochloride is a DNA-specific probe with blue fluorescence.
      Method: For cell staining (TUNEL assay).
      1. Mice are sacrificed, and their small intestine tissues are harvested and fixed with 4% formaldehyde.
      2. The tissue sections are incubated with a TUNEL reaction mixture (1 h; 37°C; dark).
      3. The sections are stained with DAPI dihydrochloride (0.1 μg/mL).
      4. Cells are observed under an inverted fluorescence microscope (Olympus, Japan).
    • Description: DAPI dihydrochloride is a DNA-specific probe with blue fluorescence.
      Method: For cell staining.
      1. Incubate cultured cells in DAPI dihydrochloride (1 μg/mL; 5 min).
      2. Observe staining cells under a fluorescence microscope.
    • Description: DAPI dihydrochloride is a DNA-specific probe with blue fluorescence.
      Method: For cell staining.
      1. Wash cultured cells for three times with PBS, cells are mounted with medium containing DAPI dihydrochloride.
      2. Samples are imaged with an Axioskop 2 mot plus fluorescence microscope equipped with Plan Apochromat 203/0.8 NA and 403/0.95 NA objectives and an AxioCam MRc camera with AxioVision software 4.7.1 (Carl Zeiss AG, Jena, Germany).
    Referencias

    • [1]. Kapuscinski J, et al. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. 1995 Sep;70(5):220-33.  [Content Brief]

    • [1]. Kapuscinski J, et al. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. 1995 Sep;70(5):220-33.

    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 (sealed storage, away from moisture and 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
    H2O / DMSO 1 mM 2.8551 mL 14.2755 mL 28.5510 mL 71.3776 mL
    5 mM 0.5710 mL 2.8551 mL 5.7102 mL 14.2755 mL
    10 mM 0.2855 mL 1.4276 mL 2.8551 mL 7.1378 mL
    15 mM 0.1903 mL 0.9517 mL 1.9034 mL 4.7585 mL
    DMSO 20 mM 0.1428 mL 0.7138 mL 1.4276 mL 3.5689 mL
    25 mM 0.1142 mL 0.5710 mL 1.1420 mL 2.8551 mL
    30 mM 0.0952 mL 0.4759 mL 0.9517 mL 2.3793 mL
    40 mM 0.0714 mL 0.3569 mL 0.7138 mL 1.7844 mL
    50 mM 0.0571 mL 0.2855 mL 0.5710 mL 1.4276 mL
    60 mM 0.0476 mL 0.2379 mL 0.4759 mL 1.1896 mL
    80 mM 0.0357 mL 0.1784 mL 0.3569 mL 0.8922 mL
    100 mM 0.0286 mL 0.1428 mL 0.2855 mL 0.7138 mL

    * Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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    DAPI dihydrochloride Related Classifications

    • Others
    • Cell Cycle/DNA Damage
    • DNA Stain
    Help & FAQs

    Keywords:

    DAPI28718-90-34',6-Diamidino-2-phenylindoleDNA StainInhibitorinhibitorinhibit

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