Home Transmembrane Transporters ATPase inhibitor (-)-Blebbistatin For research use only.

(-)-Blebbistatin

Synonyms: (S)-(-)-Blebbistatin

(-)-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.

(-)-Blebbistatin Chemical Structure

(-)-Blebbistatin Chemical Structure

CAS No. 856925-71-8

Purity & Quality Control

(-)-Blebbistatin Related Products

Signaling Pathway

ATPase Signaling Pathways

ATPase Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Rh41 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh41 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 29435139
NB-EBc1 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells 29435139
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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
In vitro

Blebbistatin, inhibits cytokinesis, alters the smooth movement of fish keratocytes and inhibits the spontaneous blebbing of a cell line lacking filamin. Blebbistatin 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]

Blebbistatin 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). Blebbistatin potently inhibits Dictyostelium myosin II, but poorly inhibits Acanthamoeba myosin II. Blebbistatin does not inhibit representative myosin superfamily members from classes I, V, and X. [2]

Blebbistatin 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. Blebbistatin 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]
Kinase Assay 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 Research Cell lines MDCK and U2OS cells
Concentrations 25 µM
Incubation Time 2-4 h
Method

Cells were treated with vlebbistatin for 2-4 hours.
Experimental Result Images Methods Biomarkers Images PMID
Western blot talin 1 / vinculin / paxillin paxillin / pY31 paxillin / pY397FAK / FAK PY epitopes / vinculin / paxillin 20308429
Growth inhibition assay Cell viability Cell death 26733241
Glycerol/urea gel electrophoresis RLC phosphorylation RLC phosphorylation RLC phosphorylation 18701651
DIC image Traction force of a palladin KD (Palld4) cell 27353427
Immunofluorescence GCs morphology actin / NMIIA / tubulin VE-cadherin / F-actin PY epitopes / actin PY epitopes / paxillin talin 1 / FAK / β1 integrin / zyxin / vinculin / α-actinin / paxillin 25598228
In Vivo
In vivo

(-)-Blebbistatin ((S)-(-)-Blebbistatin) is a cell-permeable inhibitor for non muscle myosin II ATPase.
Animal Research Animal Models Male Sprague–Dawley rats (TBI model)
Dosages 10 mg/kg
Administration i.p.

Chemical Information & Solubility

Molecular Weight 292.33 Formula

C18H16N2O2
CAS No. 856925-71-8 SDF Download (-)-Blebbistatin SDF
Smiles CC1=CC2=C(C=C1)N=C3C(C2=O)(CCN3C4=CC=CC=C4)O
Storage (From the date of receipt) 3 years -20°C(in the dark) powder

In vitro
Batch:

DMSO : 39 mg/mL ( (133.41 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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In vivo
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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

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Tech Support

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.

Handling Instructions

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