2. Membrane Transporter/Ion Channel Autophagy
  3. Na+/K+ ATPase Autophagy
  4. Ouabain Octahydrate

Ouabain Octahydrate  (Synonyms: Acocantherine; G-Strophanthin)

Cat. No.: HY-B0542 Purity: 99.96%
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Ouabain Octahydrate is an inhibitor of Na+/K+-ATPase, used for the treatment of congestive heart failure.

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

Ouabain Octahydrate Chemical Structure

Ouabain Octahydrate Chemical Structure

CAS No. : 11018-89-6

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10 mM * 1 mL in DMSO
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Based on 18 publication(s) in Google Scholar

Ouabain Octahydrate Related Antibodies

Top Publications Citing Use of Products

    Ouabain Octahydrate purchased from MedChemExpress. Usage Cited in: Med Sci Monit. 2019 Dec 11;25:9426-9434.  [Abstract]

    Ouabain leads to DNA methylation of stemness markers through increasing intracellular Ca2+ concentration. The protein levels of stemness regulators are measured in the OS cells described in

    Ouabain Octahydrate purchased from MedChemExpress. Usage Cited in: Med Sci Monit. 2019 Dec 11;25:9426-9434.  [Abstract]

    Ouabain reduces OS cell stemness but has little effects on cell viability. Western blot analysis of the protein levels of critical stemness regulators in OS cells treated with ouabain.

    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

    • Customer Review

    Description

    Ouabain Octahydrate is an inhibitor of Na+/K+-ATPase, used for the treatment of congestive heart failure.

    In Vitro

    Ouabain (100 μM) induces NLRP3 inflammasome activation and IL-1β release in macrophages. Ouabain-induced NLRP3 inflammasome activation is mediated through K+ efflux[1]. Ouabain (3 nM) alters the expression of EMT markers in NHK and ADPKD cells, and modifies cell-cell adhesion properties in ADPKD. Moreover, ouabain enhances migration of ADPKD cells, selectively modulates tight junctions, and modulates adherens junctions in ADPKD cells in a selective manner. Ouabain also activates TGFβ-Smad3 signaling, alters TER in ADPKD cells[2]. Ouabain (25, 50 or 100 nM) treatment significantly reduces cell proliferation and viability in Raji cells in a dose-dependent manner, with IC50 of 76.48±4.03 nM. Ouabain increases the number of apoptotic cells, induces autophagy, and upregulates Beclin-1 in Raji cells[4].

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

    Ouabain Octahydrate Related Antibodies
    In Vivo

    Ouabain (3 mg/kg) significantly decreases cardiac contractile force with an enlarged LVESD when mice are primed with LPS. IL-1β deficiency attenuates ouabain-induced cardiac dysfunction and injury. IL-1β secreted by infiltrated macrophages contributes to ouabain-induced cardiac inflammation. Deficiency of NLRP3 and Casp1 attenuates ouabain-induced cardiac dysfunction and macrophage infiltration[1]. Ouabain (30 µg/kg, i.p.) modulates ABCB1 activity in thymocytes of Wistar rats and it has the same effect on Swiss mice at 300 µg/kg. After 14 days of ouabain treatment, the MAP of rats is significantly elevated[3].

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

    728.77

    Formula

    C29H60O20
    CAS No.
    Appearance

    Solid

    Color

    White to off-white

    SMILES

    OC[C@@]([C@](CC1)(C[C@@H](O[C@@](O[C@@H](C)[C@H](O)[C@H]2O)([H])[C@@H]2O)C3)O)([C@@H]3O)[C@]([C@@H](C[C@@]45C)O)([H])[C@]1([H])[C@]4(CC[C@@H]5C(CO6)=CC6=O)O.[8 H2O]
    Structure Classification
    Initial Source
    Shipping

    Room temperature in continental US; may vary elsewhere.
    Storage
    Powder -20°C 3 years
    4°C 2 years
    In solvent -80°C 1 year
    -20°C 6 months
    Solvent & Solubility
    In Vitro: 

    DMSO : ≥ 125 mg/mL (171.52 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    H2O : 10 mg/mL (13.72 mM; ultrasonic and warming and heat to 60°C)

    *"≥" means soluble, but saturation unknown.

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 1.3722 mL 6.8609 mL 13.7218 mL
    5 mM 0.2744 mL 1.3722 mL 2.7443 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, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

    * 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
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    Concentration
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    Volume
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    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 (2.85 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 (2.85 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.

    For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
    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:  PBS

      Solubility: 12.5 mg/mL (17.15 mM); Clear solution; Need ultrasonic and warming and heat to 60°C

    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).
    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: 99.96%

    COA (260 KB) LCMS (114 KB)

    Handling Instructions (2659 KB)
    References
    Cell Assay
    [4]

    Cell viability is determined using a Cell Counting Kit-8 assay. Briefly, 100 μL Raji cells (5×104/mL) are seeded in triplicate in a 96-well plate and treated with various concentrations of ouabain (400, 200, 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78 and 0.39 nM) for 48 h. Following the 48-h treatment, 10 μL CCK-8 reagent is added to each well, and the cells are incubated for an additional 3 h at 37°C. Optical density (OD) values at 450 nm are subsequently measured, and each ouabain concentration is assessed in triplicate. Raji cells cultured in medium without drug served as controls. Cell viability is calculated according to the following formula: Inhibition rate (%)=[1 − (OD450(sample) − OD450(blank))/(OD450(control) − OD450(blank))] × 100.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.
    Animal Administration
    [3]

    Mice[3]
    A total of 8 mice are used in the present study, 4 in the control group and 4 in the ouabain-treated group. Animals are maintained under standard laboratory conditions, with room temperature controlled (22°C), and subjected to 12 h light-dark cycles with ad libitum access to food and water. At 24 h subsequent to the intraperitoneal injection with 300 µg/kg of ouabain or PBS, the Swiss mice are sacrificed by barbiturate overdose (86 mg/kg intraperitoneal injection of pentobarbital). The mesenteric lymph nodes and thymi are immediately removed and softly dissociated. The remaining cells are washed in PBS and centrifuged at 200 × g. The pellet is suspended in ice-cold RPMI-1640 culture medium supplemented with 10% FBS until required for the activity assays.
    Rat[3]
    Male Wistar rats are treated with daily intraperitoneal injections of 30 µg/kg of ouabain or its vehicle, phosphate-buffered saline (PBS). A total of 20 rats are used, 12 for acute treatment (n=6 rats/group in ouabain and control groups) and 8 for chronic treatment (n=4 rats/group in ouabain and control groups). Animals are maintained under standard laboratory conditions, with room temperature controlled (22°C), and subjected to 12 h light-dark cycles with ad libitum access to food and water. Prior to the first injection at 24 h and 7 and 14 days subsequent to the injection, the rats have their blood pressure measured by a computerized tail-cuff method. The animals are sacrificed by barbiturate overdose (86 mg/kg intraperitoneal injection of pentobarbital) after 24 h (acute treatment) or 14 days (chronic treatment) of ouabain injections, and the mesenteric lymph nodes, thymi and blood are collected. Full excisions of thymi and partial excisions of mesentheric lymph nodes are performed, while blood samples are collected by caudal venous puncture prior to animals sacrifice.

    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, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    H2O / DMSO 1 mM 1.3722 mL 6.8609 mL 13.7217 mL 34.3044 mL
    5 mM 0.2744 mL 1.3722 mL 2.7443 mL 6.8609 mL
    10 mM 0.1372 mL 0.6861 mL 1.3722 mL 3.4304 mL
    DMSO 15 mM 0.0915 mL 0.4574 mL 0.9148 mL 2.2870 mL
    20 mM 0.0686 mL 0.3430 mL 0.6861 mL 1.7152 mL
    25 mM 0.0549 mL 0.2744 mL 0.5489 mL 1.3722 mL
    30 mM 0.0457 mL 0.2287 mL 0.4574 mL 1.1435 mL
    40 mM 0.0343 mL 0.1715 mL 0.3430 mL 0.8576 mL
    50 mM 0.0274 mL 0.1372 mL 0.2744 mL 0.6861 mL
    60 mM 0.0229 mL 0.1143 mL 0.2287 mL 0.5717 mL
    80 mM 0.0172 mL 0.0858 mL 0.1715 mL 0.4288 mL
    100 mM 0.0137 mL 0.0686 mL 0.1372 mL 0.3430 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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    Ouabain Octahydrate 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:

    Ouabain11018-89-6Acocantherine G-StrophanthinNa+/K+ ATPaseAutophagySodium potassium pumpInhibitorinhibitorinhibit

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