Y-27632 2, inhibitor

Y-27632 dihydrochloride

Cat. Num. AAA577605

• Purity: 99.98%
• Synonyms: Y-27632 2; Y-27632 dihydrochloride; inhibitor

• Purity/Purification: 99.98%

• Formula: C14H21N3O·2HCl
• Appearance: Solid
• Receptor: ROCK1 (p160ROCK): 140 nM (Ki, cell free); ROCK2: 300 nM (Ki, cell free)
• Solubility: DMSO: 199.8 mM; Water: 43.7 mM; ; (< 1 mg/ml refers to the product slightly soluble or insoluble)
• In vitro Activity: The inhibited potency of Y-27632 against the ROCK family is 100 times than other kinases including protein kinase C, cAMP-dependent kinase and myosin light chain kinase. Y-27632 prolongs the lag time and delays the appearance of BrdU-labeledcells in a concentration-dependent manner, delays of about 1 and 4 h are noticed in the Swiss 3T3 cells treated with 10 and 100uM Y-27632, respectively [1]. The application of a selective Rho-associated kinase (ROCK) inhibitor, Y-27632, to hES cellsmarkedly diminishes dissociation-induced apoptosis, increases cloning efficiency (from approximately 1% to approximately27%) and facilitates subcloning after gene transfer. Furthermore, dissociated hES cells treated with Y-27632 are protected fromapoptosis even in serum-free suspension (SFEB) culture and form floating aggregates [2]. Y-27632 promotes neuronaldifferentiation of adipose tissue-derived stem cells (ADSCs). Compared to 1.0 and 2.5 uM Y-27632 induced groups,percentages of neuronal-like cells achieved a peak in the 5.0 uM Y-27632 induced group [3]. Y-27632 selectively inhibitssmooth-muscle contraction by inhibiting Ca2+ sensitization. We identified the Y-27632 target as a Rho-associated proteinkinase, p160ROCK. Y-27632 consistently suppresses Rho-induced [4]
• In vivo Activity: Y-27632 significantly decreased the blood pressure in a dose-dependent manner in spontaneously hypertensive rats: a fall of 50mm Hg was still observed 7 h after administration of 30 mg/kg of Y-27632. The same dose of this compound also caused asignificant and persistent fall in blood pressure in renal hypertensive rats, as well as in deoxycorticosterone acetate (DOCA)-salthypertensive rats. On the other hand, administration of the same dose of Y-27632 caused only a slight and transient fall in bloodpressure in control Wistar rats [4]. Y-27632 (5-10 mg/kg) and fasudil 5-25 (mg/kg) diminished onset of myoclonic jerks, clonicconvulsions and tonic hindlimb extensions in mice given pentylenetetrazole [5].
• Kinase Assay: Recombinant ROCK1/2, PKN, or citron kinase is expressed in HeLa cells as Myc-tagged proteins by transfection usingLipofectamine and is precipitated from the cell lysates by the use of 9E10 monoclonal anti-Myc antibody coupled to G protein-Sepharose. Recovered immunocomplexes are incubated with various concentrations of [32P]ATP and 10 mg of histone type 2as substrates in the absence or presence of various concentrations of either Y-27632 or Y-30141 at 30°C for 30 min in a totalvolume of 30 uL of the kinase buffer containing 50 mM HEPES-NaOH, pH 7.4, 10 mM MgCl2, 5 mM MnCl2, 0.02% Briji 35, and2 mM dithiothreitol. PKCa is incubated with 5 uM [32P]ATP and 200 ug/mL histone type 2 as substrates in the absence orpresence of various concentrations of either Y-27632 or Y-30141 at 30°C for 10 min in a kinase buffer containing 50 mM Tris-HCl, pH 7.5, 0.5 mM CaCl2, 5 mM magnesium acetate, 25 ug/mL phosphatidylserine, 50 ng/mL 12-O-tetradecanoyl phorbol-13-acetate and 0.001% leupeptin in a total volume of 30 uL. Incubation is terminated by the addition of 10 uL of 43 Laemmli samplebuffer. After boiling for 5 min, the mixture is subjected to SDS-polyacrylamide gel electrophoresis on a 16% gel. The gel isstained with Coomassie Brilliant Blue and then dried. The bands corresponding to histone type 2 are excised, and theradioactivity is measured [1]
• Cell Assay: Cell AssayHeLa cells are plated at a density of 3×10^4 cells per 3.5-cm dish. The cells are cultured in DMEM containing 10% FBS in thepresence of 10 mM Thymidine for 16 h. After the cells are washed with DMEM containing 10% FBS, they are cultured for anadditional 8 h, and then 40 ng/mL of Nocodazole is added. After 11.5 h of the Nocodazole treatment, various concentrations ofY-27632 (0-300 uM) or vehicle is added and the cells are incubated for another 30 min [1]
• Animal Experiment: Animal Model: Wistar rats with spontaneous or induced hypertension; Swiss albino mice with Ehrlich ascites carcinoma
• Preparation and Storage: 2 years -80°C in solvent; 3 years -20°C powder

Structure

Related Product Information for Y-27632 2, inhibitor

Y-27632 is a selective inhibitor of ROCKs including p160ROCK (Ki: 140 nM) and ROCK2 (IC50: 800 nM).