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  3. Chlorhexidine

Chlorhexidine is a orally active cationic antimicrobial agent that targets microbial cell membranes. Chlorhexidine binds to cell membrane phospholipids non-specifically, destroys membrane structure and induces leakage of cell contents. Chlorhexidine has broad-spectrum bactericidal activity against both Gram-positive and Gram-negative bacteria. Chlorhexidine can interfere with membrane permeability, cause protein precipitation and energy metabolism disorders, such as rapid inhibition of microbial growth and induction of cell death (necrosis or apoptosis).

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Chlorhexidine

Chlorhexidine Chemical Structure

CAS No. : 55-56-1

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10 mM * 1 mL in DMSO
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Customer Review

Based on 13 publication(s) in Google Scholar

Top Publications Citing Use of Products

    Chlorhexidine purchased from MedChemExpress. Usage Cited in: Cancer Res. 2019 Feb 1;79(3):534-545.  [Abstract]

    The half-life of p53 protein is detected in GYS2 overexpressed cells by added with Chlorhexidine (CHX, 20 μg/mL) for different times.
    • Biological Activity

    • Purity & Documentation

    • References

    • Customer Review

    Description

    Chlorhexidine is a orally active cationic antimicrobial agent that targets microbial cell membranes. Chlorhexidine binds to cell membrane phospholipids non-specifically, destroys membrane structure and induces leakage of cell contents. Chlorhexidine has broad-spectrum bactericidal activity against both Gram-positive and Gram-negative bacteria. Chlorhexidine can interfere with membrane permeability, cause protein precipitation and energy metabolism disorders, such as rapid inhibition of microbial growth and induction of cell death (necrosis or apoptosis)[1][2][3].

    In Vitro

    1. Cell death assay:
    Chlorhexidine (0.000125%-0.016%; 24 h) induces cell death in L929 fibroblasts, inducing a pattern of cell necrosis and/or apoptosis, as well as cell stress. Furthermore, Chlorhexidine induces apoptosis at lower concentrations and necrosis at higher concentrations, and increases the expression of heat shock protein 70 (a marker of cell stress). Chlorhexidine may have an adverse effect on the resolution of apical periodontitis[1].
    2. Cell viability experiment:
    Chlorhexidine (0.0005%-0.5%; 30 min) causes the death of canine embryonic fibroblasts with concentration of ≥0.013%, while allows the cells to survive with concentration of ≤0.006%[2].
    3. Bacterial survival experiment:
    Chlorhexidine (0.0005%-0.5%; 30 min) has a concentration-dependent bactericidal effect on Staphylococcus aureus, and kills Staphylococcus aureus with concentration of ≥0.05%, while allows the bacteria to survive with concentration of ≤0.03%[2].

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

    Cell Viability Assay[3]

    Cell Line: Canine embryonic fibroblasts
    Concentration: 0.5%, 0.05%, 0.03%, 0.013%, 0.006%, 0.005%, 0.0005%
    Incubation Time: 30 min
    Result: Fibroblasts showed 0% survival at concentrations ≥0.013%, while concentrations ≤0.006% allowed significant survival (e.g., 74% survival at 0.0005%), indicating a concentration-dependent cytotoxic effect.
    In Vivo

    1. Paw injection toxicity test:
    Chlorhexidine (0.125%-1.0%; subcutaneous injection; single dose) causes toxic effects in the plantar space of the hind paw of Balb/c mice. Concentrations ≥0.5% induces coagulative necrosis of the epidermis, dermis and subcutaneous tissues and neutrophil-dominated inflammatory responses, while concentrations ≤0.25% induces moderate inflammation and interstitial edema[1].
    2. Lung inhalation toxicity test:
    Chlorhexidine (0.125%-1%; intratracheal instillation; single dose; 8-week observation) causes collagen fiber deposition and inflammatory cell infiltration in lung tissue in the C57BL/6J mouse model, leading to restrictive ventilation dysfunction. Transcriptome analysis shows that genes related to extracellular matrix deposition and mucus secretion (such as Muc5b and Muc5ac) are upregulated[3].
    3. Genetic toxicity test:
    Chlorhexidine (0.12% Chlorhexidine dihydrochloride, 0.5 mL; oral, twice a day for 8 days) causes DNA damage in peripheral blood and oral mucosal cells of rats, without causing chromosome breakage or loss in erythrocytes[4].

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

    Animal Model: Male Balb/c mice (4 weeks old, 20 g), paw subplantar injection model[1]
    Dosage: 0.125%, 0.25%, 0.5%, 1.0% Chlorhexidine in distilled water
    Administration: Subplantar injection into the right hind paw, single dose, observed at 24 h, 48 h, 7 days, and 14 days
    Result: At 0.5% and 1.0%, caused extensive coagulative necrosis in the epidermis, dermis, and subcutaneous tissue, associated with marked neutrophilic inflammation and edema at 24-48 h; 0.25% induced focal necrosis in few specimens and moderate inflammation in all; 0.125% caused no necrosis but moderate neutrophilic inflammation and edema.
    Epidermal healing was complete by 7-14 days, with residual mononuclear cell infiltration and fibrous tissue thickening in high-dose groups.
    Animal Model: C57BL/6J mice (6-8 weeks old), intratracheal instillation-induced lung injury model[2]
    Dosage: 0.125% (2.5 mg/kg), 0.25% (5 mg/kg), 0.5% (10 mg/kg), 1% (20 mg/kg) Chlorhexidine gluconate in RO water
    Administration: Single intratracheal instillation via MicroSprayer Aerosolizer, observed for 8 weeks
    Result: Led to dose-dependent lung tissue damage, including inflammatory cell infiltration, alveolar protein deposition, and airway epithelial cell exfoliation.
    Increased collagen fiber deposition in the alveolar septum and bronchial stroma, with significantly higher lung organ coefficients in high-dose groups.
    Decreased respiratory system compliance (Crs) and forced vital capacity (FVC), indicating restrictive ventilatory dysfunction.
    Upregulated genes related to extracellular matrix organization (e.g., P4ha3) and mucus secretion (e.g., Muc5b, Muc5ac).
    Clinical Trial
    Molecular Weight

    505.45

    Formula

    C22H30Cl2N10

    CAS No.
    Appearance

    Solid

    Color

    White to off-white

    SMILES

    N=C(NC1=CC=C(Cl)C=C1)NC(NCCCCCCNC(NC(NC2=CC=C(Cl)C=C2)=N)=N)=N

    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 : 25 mg/mL (49.46 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 1.9784 mL 9.8922 mL 19.7843 mL
    5 mM 0.3957 mL 1.9784 mL 3.9569 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)

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

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

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    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 (4.95 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 (4.95 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:

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    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Please enter your animal formula composition:
    %
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    %
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    %
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    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
    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 1.9784 mL 9.8922 mL 19.7844 mL 49.4609 mL
    5 mM 0.3957 mL 1.9784 mL 3.9569 mL 9.8922 mL
    10 mM 0.1978 mL 0.9892 mL 1.9784 mL 4.9461 mL
    15 mM 0.1319 mL 0.6595 mL 1.3190 mL 3.2974 mL
    20 mM 0.0989 mL 0.4946 mL 0.9892 mL 2.4730 mL
    25 mM 0.0791 mL 0.3957 mL 0.7914 mL 1.9784 mL
    30 mM 0.0659 mL 0.3297 mL 0.6595 mL 1.6487 mL
    40 mM 0.0495 mL 0.2473 mL 0.4946 mL 1.2365 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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    Product Name:
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