Y-27632
mRNA synthesis
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.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
Y-27632是ROCK的特异性抑制剂,对ROCK1和ROCK2的Ki值分别为0.22 μM和0.30 μM。与对其它Rho效应激酶、citron激酶、PKN以及PKCα的Ki值相比,Y-27632对ROCK的抑制具有选择性。Y-27632作用于citron激酶和PKN的Ki值比ROCK激酶至少高20倍,比PKCα高大约200倍。在HeLa细胞中,Y-27632通过与ATP竞争结合,抑制ROCK1和ROCK2激酶。另外,10 μM的Y-27632抑制Swiss 3T3细胞的应力纤维 [1]。
参考文献:
[1] 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.
- 1. Longping V Tse, Rita M Meganck, et al. "Genomewide CRISPR knockout screen identified PLAC8 as an essential factor for SADS-CoVs infection." Proc Natl Acad Sci U S A. 2022 May 3;119(18):e2118126119. PMID: 35476513
- 2. Ciaran A Shaughnessy, Pamela L Zeitlin, et al. "Net benefit of ivacaftor during prolonged tezacaftor/elexacaftor exposure in vitro." J Cyst Fibros. 2022 Mar 2;S1569-1993(22)00043-1. PMID: 35248469
- 3. Heidi J Nick, Pamela L Zeitlin, et al. "Measurements of spontaneous CFTR-mediated ion transport without acute channel activation in airway epithelial cultures after modulator exposure." J Cyst Fibros. 2022 Mar 2;S1569-1993(22)00043-1. PMID: 34799640
- 4. Nina Compera, Scott Atwell, et al. "Upscaling of pneumatic membrane valves for the integration of 3D cell cultures on chip." Lab Chip. 2021 Aug 7;21(15):2986-2996. PMID: 34143169
- 5. Mirza, Amar Naeem. "The Regulation of GLI1 in Basal Cell Carcinoma." Stanford University. 2021.
- 6. Saeed Pazhoohan, Ehsan Aref, et al. "Inhibition of Rho-kinase improves response to deep inspiration in ovalbumin-sensitized guinea pigs." Iran J Basic Med Sci. 2020 Dec;23(12):1584-1589. PMID: 33489033
- 7. Preston E Bratcher, Sangya Yadav, et al. "Effect of apical chloride concentration on the measurement of responses to CFTR modulation in airway epithelia cultured from nasal brushings." Physiol Rep. 2020 Oct;8(19):e14603. PMID: 33038073
- 8. Haiyang Yu, Shan Lu, et al. "TDP-43 and HSP70 phase separate into anisotropic, intranuclear liquid spherical annuli." bioRxiv. March 29, 2020.
- 9. Dakhama A, Al Mubarak R, et al. "Tollip Inhibits ST2 Signaling in Airway Epithelial Cells Exposed to Type 2 Cytokines and Rhinovirus." J Innate Immun. 2019 Mar 29:1-13. PMID: 30928973
- 10. Zhou N, Lee JJ, et al. "Rho Kinase Regulates Aortic Vascular Smooth Muscle Cell Stiffness Via Actin/SRF/Myocardin in Hypertension." Cell Physiol Biochem. 2017 Nov 23;44(2):701-715. PMID: 29169155
- 11. Pazhoohan S, Raoufy MR, et al. "Effect of Rho-kinase inhibition on complexity of breathing pattern in a guinea pig model of asthma." PLoS One. 2017 Oct 31;12(10):e0187249. PMID: 29088265
- 12. GUANGQI ZHOU, XINMEI LIU, et al. "Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13." ONCOLOGY LETTERS,August 3, 2017.
- 13. Wang E, Wang D, et al. "Growth of hollow cell spheroids in microbead templated chambers." Biomaterials. 2017 Oct;143:57-64. PMID: 28763630
- 14. Jones BC, Kelley LC, et al. "Dual Targeting of Mesenchymal and Amoeboid Motility Hinders Metastatic Behavior." Mol Cancer Res. 2017 Jun;15(6):670-682. PMID: 28235899
- 15. Lamborn IT, Jing H, et al. "Recurrent rhinovirus infections in a child with inherited MDA5 deficiency." J Exp Med. 2017 Jul 3;214(7):1949-1972. PMID: 28606988
- 16. Wang Y, Gunasekara DB, et al. "A microengineered collagen scaffold for generating a polarized crypt-villus architecture of human small intestinal epithelium." Biomaterials. 2017 Jun;128:44-55. PMID: 28288348
- 17. Jones BC, Kelley LC, et al. "Dual Targeting of Mesenchymal and Amoeboid Motility Hinders Metastatic Behavior." Mol Cancer Res. 2017 Feb 24. PMID: 28235899
- 18. Chan W, Akhbanbetova A, et al. "Topical delivery of a Rho-kinase inhibitor to the cornea via mucoadhesive film." Eur J Pharm Sci. 2016 Aug 25;91:256-64. PMID: 27196964
Storage | Store at -20°C |
M.Wt | 247.34 |
Cas No. | 146986-50-7 |
Formula | C14H21N3O |
Solubility | ≥24.7 mg/mL in DMSO; insoluble in Chloroform |
Chemical Name | 4-[(1R)-1-aminoethyl]-N-pyridin-4-ylcyclohexane-1-carboxamide |
SDF | Download SDF |
Canonical SMILES | CC(C1CCC(CC1)C(=O)NC2=CC=NC=C2)N |
运输条件 | 蓝冰运输或根据您的需求运输。 |
一般建议 | 不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。溶液形式一般不宜长期储存,请尽快用完。 |
激酶实验 [1]: | |
激酶实验 |
通过使用Lipofectamine在HeLa细胞中转染Myc标记的重组ROCK-1、ROCK-II、PKN或citron激酶,并通过使用9E10单克隆抗Myc抗体偶联G蛋白琼脂糖凝胶从细胞裂解物中进行沉淀。在不存在或存在各种浓度的Y-27632下,将回收的免疫复合物与各种浓度的 [32P] ATP及10 mg组蛋白2作为底物一起温育,于总体积为30 mL激酶缓冲液中30°C反应30分钟,缓冲液包含50 mM HEPES-NaOH、pH 7.4、10 mM MgCl2、5 mM MnCl2、0.02%Briji 35和2 mM二硫苏糖醇。在不存在或存在各种浓度的Y-27632下,将PKCa与5 mM [32P] ATP和200 mg/ml组蛋白2作为底物,在30°C下在总体积为30 mL的激酶缓冲液中反应30 min,缓冲液配方为50 mM Tris-HCl、pH 7.5、0.5 mM CaCl2,5 mM乙酸镁,25 mg/mL磷脂酰丝氨酸,50 ng/mL 12-O-tetradecanoylphorbol-13-acetate和0.001%亮肽素。使用43 Laemmli样品缓冲液(10 mL)终止反应。 |
细胞实验 [1]: | |
细胞系 |
HeLa细胞,Swiss 3T3细胞 |
溶解方法 |
该化合物在DMSO中的溶解度大于10 mM。若获取更高浓度的溶液,可在37°C下孵育10分钟,随后在超声波浴中摇匀。-20°C以下可储存数月 |
反应条件 |
0.3和1 μM, 30 min或24 h |
应用 |
Y-27632抑制ROCK-1和ROCK-II的激酶活性,ATP以竞争性方式逆转了这种抑制作用。在Swiss 3T3细胞中,Y-27632(10 μM)消除了应激纤维,但在该浓度下,细胞周期的G1-S转变和胞质分裂几乎不受影响。在Swiss 3T3细胞中,Y-27632细胞预处理30分钟以浓度依赖性方式防止细胞形状及肌动蛋白应激纤维发生变化,在10 μM浓度下几乎完全抑制。10和100 μM的Y-27632延长了延迟时间,并以浓度依赖性方式将BrdU标记细胞的出现延迟约1和4小时。在HeLa细胞中,30 μM Y-27632抑制胞质分裂。 |
注意事项 |
由于实验环境的不同,实际溶解度可能与理论值略有不同,请测试室内所有化合物的溶解度。 |
References: [1]. 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. |
描述 | Y-27632是ROCK1(p160ROCK)的选择性抑制剂,Ki值为140 nM。 | |||||
靶点 | ROCK1 (p160ROCK) | ROCK2 | ||||
IC50 | 140 nM(Ki) | 300 nM(Ki) |
质量控制和MSDS
- 批次: