Ouabain Chemical Structure

Ouabain Chemical Structure
Molecular Weight: 728.77

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Product Description

Biological Activity

Description Ouabain is a selective Na+/K+, -ATPase inhibitor, binds to α2 /α3 subunit with Ki of 41 nM/15 nM.
Targets α3 [1] α2 [1]
IC50 15 nM(Ki) 41 nM(Ki)
In vitro Ouabain (100 nM) inhibits ATPase activities with 25%. [1] Ouabain (0.1 μM-1.0 μM) inhibits the Na+ pump and increases stored Ca2+ in cultured rat astrocytes. High Ouabain affinity isoforms (alpha2 in astrocytes, alpha3 in neurons and myocytes) are confined to a reticular distribution within the PM that paralleled underlying endoplasmic or sarcoplasmic reticulum. [2] Ouabain (0.5-1.0 mM) increases the levels of alpha1 and beta1 mRNAs, whereas it decreases those of alpha2 and beta2 mRNAs in cultured rat astrocytes. Ouabain increases alpha1 and beta1, but not alpha2 and beta2, proteins, and that the isoforms in control and ouabain-treated cultures. The ouabain-induced increase in alpha1 mRNA is blocked by cycloheximide (10 mM), the intracellular Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetraacetoxymethyl ester (30 mM), and FK506 (1 nM) in cultured rat astrocytes. [3] Ouabain (10 μM) induces significant PMD in both cell lines (28.1% of MDCK cells and 47.9% of Ma104 cells are gated in region M1, against 11.8% and 14.6% of the respective controls), but unexpectedly this effect is more remarkable in Ma104 cells. Ouabain (10 μM) induces a sustained increase in P-Tyr in MDCK cells and GSH almost completely reverted this effect, while the effect is not significant in Ma104 cells. [4]
In vivo Ouabain (14.4 mg/kg.d s.c. intermittent) further increases total peripheral resistance (TPR) in rats with heart failure due to myocardial infarction (MI), while continuous Ouabain treatment normalized TPR in rats. Ouabain (14.4 mg/kg.d s.c. continuous) significantly improves basal and maximal CO (basal: 83 mL/min; maximal: 134 mL/min). [5]
Features A glycoside poison that binds to and inhibits the action of the Na+/K+ pump in the cell membrane.

Protocol(Only for Reference)

Kinase Assay: [1]

Binding assays All binding assays are carried out with 50-100 μg of membrane protein in 1 mL aliquots containing 50 mM Tris maleate, pH 7.2, 5 mM MgCl2, 0.1 mM sodium ortho-vanadate, and different concentrations of ouabain (0.1 nM to 1 μM, specific activity ranging from 15.4 Ci/mmol to 0.12 Ci/mmol) for 2 hour at 37℃. Binding is terminated by dilution with 4 mL of ice-cold wash buffer (5 mM MgCl2, 5 mM Tris phosphate, and 50 mM Tris maleate, pH 7.4) and filtration through Whatman GF/F filters that are presoaked in 0.1% bovine serum albumin. The filters are washed four times for 2s each with 4 mL of wash buffer. Nonspecific binding, defined as the residual radioactivity seen in the presence of 2 mM unlabeled ouabain, accounted for 5-15% of total binding.

Animal Study: [5]

Animal Models Male Wistar rats with heart failure due to myocardial infarction (MI)
Formulation Saline
Dosages 14.4 mg/kg
Administration Subcutaneous

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)
Body Surface Area (m2)0.0070.0250.
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)


[1] Lucchesi PA, et al. J Biol Chem, 1991, 266(14), 9327-9331.

[2] Juhaszova M, et al. Proc Natl Acad Sci U S A, 1997, 94(5), 1800-1805.

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Chemical Information

Download Ouabain SDF
Molecular Weight (MW) 728.77


CAS No. 630-60-4
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms NSC 25485
Solubility (25°C) * In vitro DMSO 100 mg/mL (137.21 mM)
Ethanol 100 mg/mL (137.21 mM)
Water <1 mg/mL (<1 mM)
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name Card-20(22)-enolide, 3-[(6-deoxy-α-L-mannopyranosyl)oxy]-1,5,11,14,19-pentahydroxy-, hydrate (1:8), (1β,3β,5β,11α)-

Tech Support & FAQs

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
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