(-)-Blebbistatin

Catalog No.S7099 Batch:S709906

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Technical Data

Formula

C18H16N2O2
Molecular Weight 292.33 CAS No. 856925-71-8
Solubility (25°C)* In vitro DMSO 58 mg/mL (198.4 mM)
Water Insoluble
Ethanol Insoluble
In vivo (Add solvents to the product individually and in order)
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
* <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.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description (-)-Blebbistatin ((S)-(-)-Blebbistatin) is a cell-permeable inhibitor for non muscle myosin II ATPase with IC50 of ~2 μM in cell-free assays, does not inhibit myosin light chain kinase, inhibits contraction of the cleavage furrow without disrupting mitosis or contractile ring assembly.
Targets
non-muscle myosin II ATPases [2]
(Cell-free assay)
0.5 μM-5 μM
In vitro

(-)-Blebbistatin, inhibits cytokinesis, alters the smooth movement of fish keratocytes and inhibits the spontaneous blebbing of a cell line lacking filamin. This compound is shown to inhibit the enzymatic activities of HMM fragments of nonmuscle myosin IIA, nonmuscle myosin IIB and of rabbit skeletal muscle myosin S1, but not smooth muscle myosin[1]

This chemical rapidly and reversibly inhibits Mg-ATPase activity and in vitro motility of non-muscle myosin IIA and IIB for several species, while poorly inhibiting smooth muscle myosin (IC50 = 80 μM). It potently inhibits Dictyostelium myosin II, but poorly inhibits Acanthamoeba myosin II. This compound does not inhibit representative myosin superfamily members from classes I, V, and X. [2]

It does not compete with nucleotide binding to the skeletal muscle myosin subfragment-1. The inhibitor preferentially binds to the ATPase intermediate with ADP and phosphate bound at the active site, and it slows down phosphate release. This chemical interferes neither with binding of myosin to actin nor with ATP-induced actomyosin dissociation. Instead, it blocks the myosin heads in a products complex with low actin affinity. [3]
In vivo

(-)-Blebbistatin ((S)-(-)-Blebbistatin) is a cell-permeable inhibitor for non muscle myosin II ATPase.

Protocol (from reference)

Kinase Assay:

[2]
  • Actin-activated MgATPase assay

    Actin-activated MgATPase activity is measured using an NADH-coupled assay in a Beckman DU 640 spectrophotometer. (-)-Blebbistatin [a racemic mixture of the (+) and (-) enantiomers] is added from stocks dissolved in DMSO and the DMSO concentration is maintained at a constant concentration of 5% in all samples.
Cell Assay:

[4]
  • Cell lines

    MDCK and U2OS cells

  • Concentrations

    25 µM

  • Incubation Time

    2-4 h

  • Method

    Cells were treated with (-)-Blebbistatin for 2-4 hours.
Animal Study:

[5]
  • Animal Models

    Male Sprague–Dawley rats (TBI model)

  • Dosages

    10 mg/kg

  • Administration

    i.p.

References

  • https://pubmed.ncbi.nlm.nih.gov/12637748/
  • https://pubmed.ncbi.nlm.nih.gov/15548862/
  • https://pubmed.ncbi.nlm.nih.gov/15205456/
  • https://pubmed.ncbi.nlm.nih.gov/34493720/
  • https://pubmed.ncbi.nlm.nih.gov/34900532/

Customer Product Validation

(C) MyoIIA inhibition by blebbistatin treatment in vivo impairs leukemia cell extravasation into the brain. One hour before B-ALL cell transfer, C57BL/6 CD45.1+ recipient mice were treated with vehicle or blebbistatin (2.5 mg/kg). Twenty-four hours after B-ALL cell transfer, the recipient mice were intravenously injected with CD19-APC antibodies and then euthanized 4 min later.

Data from [ , , J Leukoc Biol, 2016, 143-53 ]

(a) Y27632 (20 μM) and Blebbistatin (20 μM) suppressed the nuclear translocation of YAP in HPDLCs after cyclic stretching. Scale bars indicate 50 μm.

Data from [ , , Biomed Res Int, 2018, 2018:2174824 ]

b Densitometric quantification and representative immunoblot of TCL and CM from RASMC preincubated with blebbistatin (1.25-10 μmol/l) for 1 h before exposure to 20 μg/ml ox-LDL for 24 h. Results are presented as mean ± SEM of 3 independent experiments performed in triplicate. * p < 0.05 compared with ox-LDL alone.

Data from [ , , J Vasc Res, 2016, 53(3-4):206-215 ]

Selleck's (-)-Blebbistatin Has Been Cited by 67 Publications

Surface mechanics and compressive stress impact mammalian follicle development [ Nat Commun, 2025, 16(1):9578] PubMed: 41173887
Serpina3k lactylation protects from cardiac ischemia reperfusion injury [ Nat Commun, 2025, 16(1):1012] PubMed: 39856050
Breaking the link between morphology and potency for mESCs [ Cell Biosci, 2025, 15(1):146] PubMed: 41131638
Aurora B maintains spherical shape of mitotic cells via simultaneously stabilizing myosin II and vimentin [ J Mol Cell Biol, 2025, mjaf023] PubMed: 40795355
SFTSV utilizes AXL/GAS6 for entry via PI3K-PLC-dependent macropinocytosis activated by AXL-kinase [ J Virol, 2025, 99(9):e0022125] PubMed: 40853130
Generation of two iPSC lines from patients with Aicardi-Goutières syndrome carrying either biallelic ADAR1 mutations (PC138) or a heterozygous IFIH1 mutation (PC139) [ Stem Cell Res, 2025, 89:103873] PubMed: 41351966
Molecular Control of Non-Muscle Myosin II-A Aggregation and Intracellular Dynamics by motor- or tail-specific MYH9 Mutations [ bioRxiv, 2025, 2025.05.20.654665] PubMed: 40475454
Canonical and non-canonical integrin-based adhesions dynamically interconvert [ Nat Commun, 2024, 15(1):2093] PubMed: 38453931
BMP suppresses Wnt signaling via the Bcl11b-regulated NuRD complex to maintain intestinal stem cells [ EMBO J, 2024, ] PubMed: 39433900
Methylcellulose improves dissociation quality of adult human primary cardiomyocytes [ Heliyon, 2024, 10(11):e31653] PubMed: 38841456

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.