Y-27632 dihydrochloride

Name: Y-27632 dihydrochloride
Brand: Tocris Bioscience
SKU: 1254
Rating: 5 (15 reviews)

Tocris Bioscience, part of Bio-Techne | Catalog # 1254

(15)

Citations (1537)

COA / SDS

Selective ROCK inhibitor; inhibits ROCK1 (p160 ROCK) and ROCK2; stem cell freezing (cryopreservation) media component

Key Product Details

Description:	Selective ROCK inhibitor; inhibits ROCK1 (p160 ROCK) and ROCK2; stem cell freezing (cryopreservation) media component
Alternative Names:	y27632,Y-27632 2HCl,ROCK inhibitor,ROCK1 inhibitor
Chemical Name:	trans-4-[(1R)-1-Aminoethyl]-N-4-pyridinylcyclohexanecarboxamide dihydrochloride
Purity:	≥98% (HPLC)
Molecular Weight:	320.26

View Full Technical Details for Y-27632 dihydrochloride

Technical Data
Citations / Reviews (15)

Biological Activity for Y-27632 dihydrochloride

Y-27632 dihydrochloride is a selective ROCK inhibitor (K_i values are 0.14-0.22, 0.3, 25, 26 and > 250 μM for ROCK1 (p160 ROCK), ROCK2, PKA, PKC and MLCK respectively). Also inhibits PRK2 (IC₅₀ = 600 nM). Increases survival rate of human embryonic stem (hES) cells and iPSC undergoing cryopreservation. Used in combination with CHIR 99021, RepSox, Forskolin, SP 600125, Go 6983 and VPA to reprogram fibroblasts to mature neurons. Also optimizes naive human pluripotent stem cell growth and viability following naive cell derivation from primed ESCs and iPSCs using naive human stem cell medium (NHSM). Used as component of growth media for urothelial organoids. Also used in a protocol to generate brain organoids from human iPSCs. Cell-permeable, orally bioavailable and active in vivo.

Y-27632 in solution and Y-27632 synthesized to cGMP guidelines also available.

For more information about how Y-27632 dihydrochloride may be used, see our protocols: Transdifferentiating Fibroblasts into Neurons, Converting Fibroblasts into Cardiomyocytes (9C Cocktail), Accelerated Induction of Cortical Neurons from hiPSCs, Human iPSC-derived Myoblasts, Generation and Propagation of EPS cells (LCDM Cocktail), Generating Vascular Organoids, Generating Midbrain Dopaminergic Neurons from hPSCs, Cultivating Cerebral Organoids, 3D Culture of Lung Alveolar Cells.

Compound Libraries for Y-27632 dihydrochloride

Y-27632 dihydrochloride is also offered as part of the Tocriscreen 2.0 Max, Tocriscreen Kinase Inhibitor Library and Tocriscreen Stem Cell Library. Find out more about compound libraries available from Tocris.

Scientific Data Examples for Y-27632 dihydrochloride

	Application of Y-27632 dihydrochloride in MESP1+ Mesoderm. ROCK inhibition by Y-27632 (1 μM) promotes skeletal myogenic differentiation of MESP1+ mesoderm, as shown by immunostaining for Myosin Heavy Chain+ (Green) and Myogenin (Red), and counterstaining with DAPI (Catalog # 5748; Blue, Tocris). Data courtesy of Dr Sunny Chan and Dr Michael Kyba, University of Minnesota.
	Application of Y-27632 dihydrochloride in Culture and Characterization of iPSC-derived Cerebral Organoids. iPSC-derived cerebral organoids (day 45) were cultured using Cultrex UltiMatrix RGF Basement Membrane Extract (Catalog # BME001-05) and brain organoid culture medium, which includes Y-27632 dihydrochloride, along with the other reagents listed in the brain organoid culture recipe. Cerebral organoids were stained for Syto6 (blue), Pax6 (red), and Vimentin (green). (A) Image taken at 4x magnification. (B) An enlarged view of the area shown within the white box in part A of the figure. (C) Image taken at 15x magnification. Images courtesy of LifeCanvas Technologies.
	Application of Y-27632 dihydrochloride in Culture and Characterization of iPSC-derived Cerebral Organoids. iPSC-derived cerebral organoids (day 45) were cultured using Cultrex UltiMatrix RGF Basement Membrane Extract (Catalog # BME001-05) and brain organoid culture medium, which includes Y-27632 dihydrochloride, along with the other reagents listed in the brain organoid culture recipe. (A) Representative brightfield image of day 30 iPSC-derived cerebral organoids. (B) Cerebral organoids stained for beta III-tubulin (green) and Prox1 (red), and counterstained with DAPI (blue) (Catalog # 5748).
	Application of Y-27632 dihydrochloride in Culture and Characterization of iPSC-derived Human Intestinal Organoids. iPSC-derived human intestinal organoids were cultured using Y-27632 dihydrochloride, SB 202190 (Catalog # 1264), N-Acetylcysteine (Catalog # 7874), A 83-01 (Catalog # 2939), Nicotinamide (Catalog # 4106), along with the other reagents used in the human intestinal organoid culture protocol. (A) Human intestinal organoids were stained using a Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (Catalog # MAB2105; green) and a Goat Anti-Human/Mouse Desmin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3844; red) to visualize myofibroblast cells and counterstained with DAPI (Catalog # 5748; blue). (B) Human intestinal organoids were stained using a Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF748; green), a Mouse Anti-Human MUC2 Monoclonal Antibody (Novus Biologicals, Catalog # NBP2-44431; red) and counterstained with DAPI (Catalog # 5748; blue).
	Application of Y-27632 dihydrochloride in Culture and Characterization of Adult Stem Cell-derived Liver Organoids. Adult stem cell-derived liver organoids were cultured using Y-27632 dihydrochloride (Catalog # 1254), HEPES (Catalog # 3173), A 83-0 (Catalog # 2939), Forskolin (Catalog # 1099), N-Acetylcysteine (Catalog # 7874), along with the other reagents used in the human liver organoid culture protocol. After 3 days, liver organoid expansion medium was added for 7-10 days, followed by liver organoid differentiation medium, which includes HEPES (Catalog # 3173), Nicotinamide (Catalog # 4106), N-Acetylcysteine (Catalog # 7874), A 83-0 (Catalog # 2939), along with the other reagents used in the human liver organoid culture protocol. Differentiated human liver organoids were stained using a (A) Mouse Anti-Human Serum Albumin Monoclonal Antibody (Catalog # MAB1455; red) and a (B) Goat Anti-Human HNF-3beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400; red) and counterstained with DAPI (Catalog # 5748; blue).
	Application of Y-27632 dihydrochloride in Culture and Characterization of Adult Stem Cell-derived Human Descending Colon Organoids. Adult stem cells isolated from human descending colon were embedded in Cultrex UltiMatrix RGF Basement Membrane Extract (Catalog # BME001-05) and cultured for 30 days in intestinal organoid culture medium, which includes Y-27632 dihydrochloride, SB 202190 (Catalog # 1264), N-Acetylcysteine (Catalog # 7874), A 83-01 (Catalog # 2939), Nicotinamide (Catalog # 4106),, along with the other reagents used in the human intestinal organoid culture protocol. (A) Organoids were fixed and stained with a Mouse Anti-Human MUC2 Monoclonal Antibody (Novus Biologicals; Catalog # NBP2-44431; green) to visualize intestinal goblet cells and counterstained with a Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3844; red) and DAPI (Catalog # 5748; blue). The image shown was taken at 10x magnification. (B) Organoids were fixed and stained with a Mouse Anti-Human Chromogranin A Monoclonal Antibody (Catalog # MAB90981; green) to visualize enteroendocrine cells and counterstained with a Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3844; red) and DAPI (Catalog # 5748; blue). The image shown was taken at 20x magnification.
	Application of Y-27632 dihydrochloride in Culture and Characterization of Adult Stem Cell-derived Lung Organoids. Adult stem cells isolated from human lung biopsy tissue were embedded in Cultrex UltiMatrix RGF Basement Membrane Extract (Catalog # BME001-05) and cultured for 20-60 days in lung organoid expansion medium, which includes Y-27632 dihydrochloride, HEPES (Catalog # 3173), N-Acetylcysteine (Catalog # 7874), A 83-01 (Catalog # 2939), SB 202190 (Catalog # 1264), Nicotinamide (Catalog # 4106), along with the other reagents used in the human lung organoid culture protocol. Lung organoids were able to differentiate and exhibit markers for various cell types of the lung. Lung organoids were stained with (A) a Rabbit Anti-Human Cytokeratin 5 (KRT5) Monoclonal Antibody (green) and a Goat Anti-Human p63/TP73L Polyclonal Antibody (Catalog # AF1916; red) to visualize basal cells, (B) a Hamster Anti-Mouse Podoplanin (PDPN) Monoclonal Antibody (Novus Biologicals, Catalog # NB600-1015; green) to visualize alveolar type I cells and a Goat Anti-Human p63/TP73L Polyclonal Antibody (Catalog # AF1916; red) to visualize basal cells, and (C, D) a Mouse Anti-MUC5AC Monoclonal Antibody (Novus Biologicals, Catalog # NBP2-15196; green) to visualize Goblet cells and a Mouse Anti-Human/Mouse/Rat SOX2 Monoclonal Antibody (Catalog # MAB2018; red). All samples were counterstained with DAPI (Catalog # 5748; blue).

Technical Data for Y-27632 dihydrochloride

M. Wt	320.26
Formula	C₁₄H₂₁N₃O.2HCl
Storage	Desiccate at RT
Purity	≥98% (HPLC)
CAS Number	129830-38-2
PubChem ID	14801906
InChi Key	IDDDVXIUIXWAGJ-LJDSMOQUSA-N
Smiles	N[C@H](C)[C@@]1([H])CC[C@H](C(NC2=CC=NC=C2)=O)CC1.Cl.Cl

The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.

Tocris products are intended for laboratory research use only, unless stated otherwise.

Solubility for Y-27632 dihydrochloride

	Solvent	Max Conc. mg/mL	Max Conc. mM
Solubility
	water	32.03	100
	DMSO	32.03	100

Preparing Stock Solutions for Y-27632 dihydrochloride

The following data is based on the product molecular weight 320.26.

Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.

Select a batch to recalculate based on the batch molecular weight:

Concentration / Solvent Volume / Mass	1 mg	5 mg	10 mg
1 mM	3.12 mL	15.61 mL	31.22 mL
5 mM	0.62 mL	3.12 mL	6.24 mL
10 mM	0.31 mL	1.56 mL	3.12 mL
50 mM	0.06 mL	0.31 mL	0.62 mL

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References for Y-27632 dihydrochloride

References are publications that support the biological activity of the product.

Schafer An in vivo neuroimmune organoid model to study human microglia phenotypes. Cell 2023 PMID: 37172564
Chan et al Development of bipotent cardiac/skeletal myogenic progenitors from MESP1+ mesoderm. Stem Cell Rep. 2016 PMID: 26771351
Santos Urothelial organoids originating from Cd49f^high mouse stem cells display Notch-dependent differentiation capacity. Nat.Commun. 2019 PMID: 31562298
Ishizaki Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases. Mol.Pharmacol. 2000 PMID: 10779382
Hu Direct conversion of normal and Alzheimer's Disease human fibroblasts into neuronal cells by small molecules. Cell Stem Cell 2015 PMID: 26253202
Jung Isolation and in vitro expansion of human colonic stem cells. Nat.Med. 2011 PMID: 21892181
Wimmer Human blood vessel organoids as a model of diabetic vasculopathy. Nature 2019 PMID: 30651639
Bartfeld In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection. Gastroenterology 2015 PMID: 25307862
Lancaster Generation of Cerebral Organoids from Human Pluripotent Stem Cells Nat. Protoc. 2015 PMID: 25188634
Sato SnapShot: Growing Organoids from Stem Cells. Cell 2015 PMID: 26091044
Mahe Establishment of gastrointestinal epithelial organoids Curr.Protoc.Mouse.Biol 2014 PMID: 25105065
Wilson Cryopreservation of brain endothelial cells derived from human induced pluripotent stem cells is enhanced by Rho-associated coiled coil-containing kinase inhibition. Tissue.Eng.Part C.Methods 2016 PMID: 27846787
Gafni Derivation of novel human ground state naive pluripotent stem cells. Nature 2013 PMID: 24172903
Ichikawa Freeze-thawing single human embryonic stem cells induce e-cadherin and actin filament network disruption via g13 signaling. Cryo Letters. 2011 PMID: 22227712
Uehata Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase. Nature 1997 PMID: 9353125
Narumiya Use and properties of ROCK-specific inhibitor Y-27632. Methods Enzymol. 2000 PMID: 11036610