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> Propidium (Iodide) [25535-16-4]

Propidium (Iodide) [25535-16-4]

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Cat# HY-D0815-50mg

Size : 50mg

Brand : MedChemExpress

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Propidium Iodide (PI) is a nuclear staining agent that stains DNA. Propidium Iodide is an analogue of ethidine bromide that emits red fluorescence upon embedding in double-stranded DNA. Propidium Iodide cannot pass through living cell membranes, but it can pass through damaged cell membranes to stain the nucleus. Propidium Iodide has a fluorescence wavelength of 493/617 nm and a wavelength of 536/635 nm after Mosaic with DNA. Propidium Iodide is commonly used in the detection of apoptosis (apoptosis) or necrosis (necrosis), and is often used in flow cytometry analysis.

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

Propidium Iodide Chemical Structure

Propidium Iodide Chemical Structure

CAS No. : 25535-16-4

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

Based on 59 publication(s) in Google Scholar

    Propidium Iodide purchased from MedChemExpress. Usage Cited in: ACS Nano. 2023 Feb 8.  [Abstract]

    Propidium iodide (PI) staining for cell death with GCFN and ferroptosis inhibitor ferrostatin-1 treatment for 48 h

    Propidium Iodide purchased from MedChemExpress. Usage Cited in: Eur J Pharmacol. 2022 Apr 5;920:174830.  [Abstract]

    Primary newborn rat cardiac fibroblasts are stained with PI (2 μg/mL; for 2 h) and analyzed using a Nikon Ti2 microscope.

    Propidium Iodide purchased from MedChemExpress. Usage Cited in: Molecules. 2022 Oct 9;27(19):6733.  [Abstract]

    In A549 cells, Propidium iodide (PI, 50 μg/mL) is added for incubation for 30 min in the dark. Cell-cycle distribution is determined using a FACSaria-I flow cytometer.

    Propidium Iodide purchased from MedChemExpress. Usage Cited in: Thorac Cancer. 2022 May;13(9):1299-1310.  [Abstract]

    KYSE450 and ECA109 cells after 96 h various transfections were washed three times in cold PBS, stained with Propidium iodide (PI, 50 μg/mL) and Annexin V‐FITC (25 μg/mL) for 20 min in the dark, and analyzed within 1 h.

    Propidium Iodide purchased from MedChemExpress. Usage Cited in: Redox Biol. 2022 Feb;49:102207.  [Abstract]

    PAMs and PK-15CD163 cells sre washed with phosphate-buffered saline (PBS), and then incubated with the nuclear dye propidium iodide for 15 min. The nuclei sre counterstained with propidium iodide (red). Fluorescent images are acquired with a confocal laser scanning microscope.

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

    The NP tissue slices are incubated with a culture medium containing 1 μg/ml Calcein AM and 1 μg/ml PI for one hour at 37°C. After washing with PBS, the slices are immediately viewed using an inverted confocal laser scanning microscope.

    Propidium Iodide purchased from MedChemExpress. Usage Cited in: Aging (Albany NY). 2018 Nov 28;10(11):3353-3370.  [Abstract]

    KLE cells are stained by 25 μg/mL PI in fluorescence-activated cell sorting buffer, incubated cells for 30 min in dark environment at room temperature. Cell apoptosis is determined by Annexin V/PI double staining and percentages of cells in each phase are calculated.

    Propidium Iodide 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.
    Description

    Propidium Iodide (PI) is a nuclear staining agent that stains DNA. Propidium Iodide is an analogue of ethidine bromide that emits red fluorescence upon embedding in double-stranded DNA. Propidium Iodide cannot pass through living cell membranes, but it can pass through damaged cell membranes to stain the nucleus. Propidium Iodide has a fluorescence wavelength of 493/617 nm and a wavelength of 536/635 nm after Mosaic with DNA. Propidium Iodide is commonly used in the detection of apoptosis (apoptosis) or necrosis (necrosis), and is often used in flow cytometry analysis.

    In Vitro

    "General Protocol

    1.Preparation of PI working solution
    1.1 Preparation of the stock solution
    Dissolve 1 mg PI 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 PI working solution
    Dilute the stock solution in serum-free cell culture medium or PBS to obtain 20-50 μg/mL of working solution.
    Note: Please adjust the concentration of PI 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 5-10 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 5-10 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 PI 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.

    Propidium Iodide Related Antibodies
    Molecular Weight

    668.39

    Formula

    C27H34I2N4
    CAS No.

    25535-16-4
    Appearance

    Solid

    Color

    Pink to red

    Emission (Em)

    615
    Excitation (Ex)

    535
    SMILES

    C[N+](CCC[N+]1=C(C2=CC=CC=C2)C3=CC(N)=CC=C3C4=C1C=C(N)C=C4)(CC)CC.[I-].[I-]
    Shipping

    Room temperature in continental US; may vary elsewhere.
    Storage

    4°C, sealed storage, away from moisture and light

    *In solvent : -80°C, 2 years; -20°C, 1 year (sealed storage, away from moisture and light)

    Solvent & Solubility
    In Vitro: 

    DMSO : 100 mg/mL (149.61 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    H2O : 3.57 mg/mL (5.34 mM; ultrasonic and warming and heat to 60°C)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 1.4961 mL 7.4807 mL 14.9613 mL
    5 mM 0.2992 mL 1.4961 mL 2.9923 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, 2 years; -20°C, 1 year (sealed storage, away from moisture and light). When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

    * 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.

    • 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 (3.74 mM); Clear solution

      This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

      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 (3.74 mM); Clear solution

      This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

      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, 2 years; -20°C, 1 year (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.
    Purity & Documentation

    Purity: 99.69%

    COA (249 KB) HNMR (261 KB) RP-HPLC (244 KB) LCMS (94 KB)

    Handling Instructions (2659 KB)
    Dyeing Example
    • Description: Propidium Iodide is a cells dye with red fluorescence. Propidium Iodide only can be taken up by cells with impaired membrane integrity, it can be used in cell cycle and cell viability assays.
      Method: For cell nucleus staining.
      1. Cells are washed with PBS, and then incubates cells with Propidium Iodide (15 min).
      2. Fluorescent images are acquired with a confocal laser scanning microscope (Olympus Fluoviewer. 3.1, Japan).
    • Description: Propidium Iodide is a cells dye with red fluorescence. Propidium Iodide only can be taken up by cells with impaired membrane integrity, it can be used in cell cycle, cell viability and necrocytosis assays.
      Method: For cell walls staining.
    • Description: Propidium Iodide is a cells dye with red fluorescence. Propidium Iodide only can be taken up by cells with impaired membrane integrity, it can be used in cell cycle, cell viability and necrocytosis assays.
      Method: For cell staining.
      1. Fix cultured cells with paraformaldehyde (4%; 10 min).
      2. Incubate cells with Propidium Iodide (50 mg/L; 15 min; dark).
      3. Fluorescent images are acquired with a confocal laser scanning microscope.
    References

    • [1]. Hezel M, et al. Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain. Micron. 2012 Oct;43(10):1031-8.  [Content Brief]

      [2]. Nicoletti I, et al. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9.  [Content Brief]

      [3]. A rapid and simple method for measuring thymocyte apoptosis by propidium iodidestaining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9.  [Content Brief]

    Cell Assay
    [2]

    Flow cytometric analysis: Propidium iodide is prepared in in 0.1% sodium citrate plus 0.1% Triton X-100 (50 μg/mL). The 200 ×g centrifuged cell pellet is gently resuspended in 1.5 mL hypotonlc fluorochrome solution (Propidium iodide 50 μg/mL), in 12×75 polypropylene tubes. The tubes are placed at 4°C in the dark overnight before the flow cytometric analysis. The propidium Iodide fluorescence of individual nuclei is measured using a FACScan flow cytometer. The nuclei traverses the light beam of a 488 nm Argon laser. A 560 nm dichrolc nurror (DM 570) and a 600 nm band pass filter (bandwidth 35 nm) are used for collecting the red fluorescence due to propidium Iodide staining of DNA and the data are registered on a logarithmic scale[2].

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

    • [1]. Hezel M, et al. Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain. Micron. 2012 Oct;43(10):1031-8.  [Content Brief]

      [2]. Nicoletti I, et al. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9.  [Content Brief]

      [3]. A rapid and simple method for measuring thymocyte apoptosis by propidium iodidestaining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9.  [Content Brief]

    • [1]. Hezel M, et al. Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain. Micron. 2012 Oct;43(10):1031-8.

      [2]. Nicoletti I, et al. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9.

      [3]. A rapid and simple method for measuring thymocyte apoptosis by propidium iodidestaining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9.

    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, 2 years; -20°C, 1 year (sealed storage, away from moisture and light). When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    H2O / DMSO 1 mM 1.4961 mL 7.4807 mL 14.9613 mL 37.4033 mL
    5 mM 0.2992 mL 1.4961 mL 2.9923 mL 7.4807 mL
    DMSO 10 mM 0.1496 mL 0.7481 mL 1.4961 mL 3.7403 mL
    15 mM 0.0997 mL 0.4987 mL 0.9974 mL 2.4936 mL
    20 mM 0.0748 mL 0.3740 mL 0.7481 mL 1.8702 mL
    25 mM 0.0598 mL 0.2992 mL 0.5985 mL 1.4961 mL
    30 mM 0.0499 mL 0.2494 mL 0.4987 mL 1.2468 mL
    40 mM 0.0374 mL 0.1870 mL 0.3740 mL 0.9351 mL
    50 mM 0.0299 mL 0.1496 mL 0.2992 mL 0.7481 mL
    60 mM 0.0249 mL 0.1247 mL 0.2494 mL 0.6234 mL
    80 mM 0.0187 mL 0.0935 mL 0.1870 mL 0.4675 mL
    100 mM 0.0150 mL 0.0748 mL 0.1496 mL 0.3740 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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    Propidium Iodide Related Classifications

    • Others
    • Others Cell Cycle/DNA Damage
    • Fluorescent Dye DNA/RNA Synthesis
    Help & FAQs

    Keywords:

    Propidium25535-16-4Fluorescent DyeDNA/RNA SynthesisInhibitorinhibitorinhibit

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