In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
Y-27632 is a specific inhibitor of Rho-associated kinases (ROCK) family with Ki values of 0.22 μM and 0.30 μM for ROCK1 and ROCK2, respectively.
Y-27632 has shown selectivity of inhibition by comparing their Ki values for other Rho effector kinases, citron kinase and PKN, as well as PKCα. The Ki values of Y-27632 for citron kinase and PKN are least 20-fold higher, and Ki values for PKCα are about 200-fold higher than those for ROCK kinases. In addition, Y-27632 has been reported to inhibit ROCK1 and ROCK2 by competing with ATP for binding to the kinase in HeLa cells. Besides, Y-27632 has also shown the inhibition of stress fibers in Swiss 3T3 cells when the concentration of Y-27632 is 10 μM .
 Ishizaki T1, Uehata M, Tamechika I, Keel J, Nonomura K, Maekawa M, Narumiya S. Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases. Mol Pharmacol. 2000 May;57(5):976-83.
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|Storage||Store at -20°C|
|Solubility||≥24.7 mg/mL in DMSO; insoluble in Chloroform|
|Shipping Condition||Evaluation sample solution: ship with blue ice. All other available sizes: ship with RT, or blue ice upon request.|
|General tips||For obtaining a higher solubility, please warm the tube at 37°C and shake it in the ultrasonic bath for a while. Stock solution can be stored below -20°C for several months.|
|Kinase experiment :|
Recombinant ROCK-I, ROCK-II, PKN, or citron kinase was expressed in HeLa cells as Myc-tagged proteins by transfection using Lipofectamine, and was precipitated from the cell lysates by the use of 9E10 monoclonal anti-Myc antibody coupled to G protein-Sepharose. Recovered immunocomplexes were incubated with various concentrations of [32P] ATP and 10 mg of histone type 2 as substrates in the absence or presence of various concentrations of either Y-27632 at 30°C for 30 min in a total volume of 30 mL of the kinase buffer (50 mM HEPES-NaOH, pH 7.4, 10 mM MgCl2, 5 mM MnCl2, 0.02% Briji 35, and 2 mM dithiothreitol). PKCa was incubated with 5 mM [32P] ATP and 200 mg/mL histone type 2 as substrates in the absence or presence of various concentrations of either Y-27632at 30°C for 10 min in a kinase buffer (50 mM Tris-HCl, pH 7.5, 0.5 mM CaCl2, 5 mM magnesium acetate, 25 mg/mL phosphatidyl serine, 50 ng/mL 12-O-tetradecanoylphorbol-13-acetate and 0.001% leupeptin) in a total volume of 30 mL. Incubation was terminated by 43 Laemmli sample buffer (10 mL).
|Cell experiment :|
HeLa cells, Swiss 3T3 cells
The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37°C for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20°C for several months.
0.3 and 1 μM, 30 min or 24 h
Y-27632 inhibited the kinase activity of both ROCK-I and ROCK-II in vitro, and this inhibition was reversed by ATP in a competitive manner. Y-27632 abolished stress fibers in Swiss 3T3 cells at 10 μM, but the G1-S phase transition of the cell cycle and cytokinesis were little affected at this concentration. In Swiss 3T3 cells, prior treatment of cells with Y-27632 for 30 min prevented cell shape and actin-stress fibers changes in a concentration-dependent manner; almost complete inhibition was observed at 10 μM. 10 and 100 μM Y-27632 prolonged the lag time and delayed the appearance of BrdU-labeled cells in a concentration-dependent manner, delays of about 1 and 4 h. In HeLa cells, 30 μM Y-27632 inhibited cytokinesis.
Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.
. Ishizaki T, Uehata M, Tamechika I, et al. Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases[J]. Molecular pharmacology, 2000, 57(5): 976-983.
|Description||Y-27632 2HCl is a selective inhibitor of ROCK1 (p160ROCK) with a Ki value of 140 nM.|
|IC50||140 nM(Ki)||300 nM(Ki)|
Quality Control & MSDS
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