Y-27632 dihydrochloride

Tocris Bioscience | Catalog # 1254

GMP Version Available: TB1254-GMP
GMP
Selective ROCK inhibitor; inhibits ROCK1 (p160 ROCK) and ROCK2; stem cell freezing (cryopreservation) media component
Tocris Bioscience

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
View all Rho-kinase Inhibitors »

Product Description

Y-27632 dihydrochloride is a selective ROCK inhibitor (Ki 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.

Scientific Data Images for Y-27632 dihydrochloride

ROCK inhibition by Y-27632 and immunostaining of skeletal myogenic differentiation of MESP1+ mesoderm,

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.

iPSC-derived cerebral organoids

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 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.

iPSC-derived cerebral organoids

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 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).

iPSC-derived Human Intestinal Organoid

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 other reagents used in the human intestinal organoid culture protocol. (A) Human intestinal organoids were stained using a Human/Mouse/Rat Vimentin Antibody (Catalog # MAB2105; green) and a Human/Mouse Desmin Antibody (Catalog # AF3844; red) to visualize myofibroblast cells and counterstained with DAPI (Catalog # 5748; blue). (B) Human intestinal organoids were stained using a Human/Mouse E-Cadherin Antibody (Catalog # AF748; green), a Human MUC2 Antibody (Novus Biologicals, Catalog # NBP2-44431; red) and counterstained with DAPI (Catalog # 5748; blue).

Adult Stem Cell-derived Liver Organoids

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, HEPES (Catalog # 3173), A 83-01 (Catalog # 2939), Forskolin (Catalog # 1099), N-Acetylcysteine (Catalog # 7874), along with 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 83-01 (Catalog # 2939), along with the other reagents. Differentiated human liver organoids were stained using a (A) Human Serum Albumin Antibody (Catalog # MAB1455; red) and a (B) Human HNF-3beta /FoxA2 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 and then 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 other reagents used in the human intestinal organoid culture protocol. (A) Organoids were fixed and stained with a Human MUC2 Antibody (Novus Biologicals; Catalog # NBP2-44431; green) to visualize intestinal goblet cells and counterstained with a Human/Mouse E-Cadherin Antibody (Catalog # AF748; red) and DAPI (Catalog # 5748; blue).(B) Organoids were fixed and stained with a Human Chromogranin A Antibody (Catalog # MAB90981; green) to visualize enteroendocrine cells and counterstained with a Human/Mouse E-Cadherin Antibody (Catalog # AF748; red) and DAPI (Catalog # 5748; blue).

Adult Stem Cell-derived Lung Organoids

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 and then 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 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 Human Cytokeratin 5 (KRT5) Antibody (green) and a Human p63/TP73L Antibody (Catalog # AF1916; red) to visualize basal cells, (B) a Mouse Podoplanin (PDPN) Antibody (Novus Biologicals, Catalog # NB600-1015; green) to visualize alveolar type I cells and a Human p63/TP73L Antibody (Catalog # AF1916; red) to visualize basal cells, and (C, D) a Mouse MUC5AC Antibody (Novus Biologicals, Catalog # NBP2-15196; green) to visualize Goblet cells and a Human/Mouse/Rat SOX2 Antibody (Catalog # MAB2018; red). All samples were counterstained with DAPI (Catalog # 5748; blue).

Product Specifications for Y-27632 dihydrochloride

Molecular Weight

320.26

Formula

C14H21N3O.2HCl

Storage

Desiccate at RT

Purity

≥98% (HPLC)

Chemical Name

trans-4-[(1R)-1-Aminoethyl]-N-4-pyridinylcyclohexanecarboxamide dihydrochloride

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.

Solubility

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 all 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

Calculators

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Background References

References are publications that support the biological activity of the product. See our Citations tab to view 1643 publications citing the usage of this product.

Product Documents for Y-27632 dihydrochloride

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Y-27632 dihydrochloride

For research use only

Citations for Y-27632 dihydrochloride

Customer Reviews for Y-27632 dihydrochloride (19)

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Customer Images

Showing  1 - 5 of 19 reviews Showing All
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  • Helps iPSC attach
    Name: Eric Wang
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: iPSC
    Verified Customer | Posted 07/11/2025
    For stem cell culture
  • Y-27632
    Name: Chi Ngo
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: human iPSC
    Verified Customer | Posted 11/25/2024
    I used the Y-27632 compound to help with iPSC's viability after thaw.
    Y-27632 dihydrochloride 1254
  • Consistent performance
    Name: Ted Mason
    Species: Human
    Assay Type: In Vitro
    Verified Customer | Posted 08/22/2023
    ROCKi has consistently performed in iPSC cell culture work. Well packaged vials are resuspended and aliquoted for -20 storage. Cells passaged or fed with ROCKi media have shown good viability. This is a regular purchase for the labs at a reasonable price. Images @10x magnification show limited differentiation of iPSCs after 3 days in culture.
    Y-27632 dihydrochloride 1254
  • Great for increasing organoids viability
    Name: MARIA PINNA
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: Organoids models
    Verified Customer | Posted 05/17/2023
    Supplementing the organoids culture media with Y-27632 dihydrochloride after passaging prevents cell death and increases viability.
  • Excellent product
    Name: Jack Schnell
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: iPSCs
    Verified Customer | Posted 03/15/2023
    Easy to manage and works as expected
    We add this product at a concentration of 10 uM to promote cell survival in in vitro iPSC cultures.
    Y-27632 dihydrochloride 1254
  • In vitro study
    Name: Emma Watson
    Species: Human
    Verified Customer | Posted 08/31/2021
    for human iPSC cells
  • PDX organoid culture
    Name: Puru Lamichhane
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: PDX
    Verified Customer | Posted 03/14/2021
    PDX organoid culture
  • ROCK Inhibitor
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: Organoids
    Verified Customer | Posted 09/07/2020
    To generate human intestinal organoids
  • test
    Name: Veselina Petrova
    Species: Human
    Assay Type: In Vitro
    Verified Customer | Posted 08/28/2020
    I used this product on freshly split and plate human iPSC cells to help cell survival and adherence. I used the product at 10 mM concentration which I diluted in 1:1000 in culture media and left the cells at 37C, 5% CO2 for 24 hours before replacing with fresh culture media.
  • For hPSC culture
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: Human PSC
    Verified Customer | Posted 06/30/2020
    For hPSC culture, final concentration 10mM. Reconstitute 10 mg of Y27632 in 3.0792 ml of PBS. Make 100µl aliquots and keep them at −20 °C for up to 6 months.
    Y-27632 dihydrochloride 1254
  • Worked great and at a great price
    Name: Tres Davis
    Species: Mouse
    Assay Type: Ex Vivo
    Verified Customer | Posted 05/08/2020
    HPLC
  • Neural Induction Conditions
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: H9
    Verified Customer | Posted 03/22/2020
    ROCK inhibitor Y-27632 (10 mM)
    Y-27632 dihydrochloride 1254
  • Great for passaging stem cells.
    Name: Samiha Shaikh
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: iPSC
    Verified Customer | Posted 01/31/2020
    Great for passaging huiPSCs with dissociation into single cells.
    Y-27632 dihydrochloride 1254
  • Great for single iPS cell survival
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: iPSC
    Verified Customer | Posted 09/13/2019
    I use at 1:1000 and store at -20C. I remove the compound from medium 24 hours after plating.
    Used during single cell passage for CRISPR clone isolation, works well and no difference found between multiple batches. Cells show characteristic 'spiky' morphology upon addition to the medium.
    Y-27632 dihydrochloride 1254
  • Human PDAC orgnaoids
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: tissue
    Verified Customer | Posted 05/03/2019
    To make Human PDAC orgnaoids
    Y-27632 dihydrochloride 1254
  • For iPSC culture
    Name: Pinyuan Tian
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: iPSC
    Verified Customer | Posted 08/07/2018
    Make 100µl aliquots and keep them at −20 °C for up to 6 months.
    For iPSC culture, final concentration 10mM. Reconstitute 10 mg of Y27632 in 3.0792 ml of PBS.
    Y-27632 dihydrochloride 1254
  • ROCK Inhibitor to improve cartilage integrity
    Name: Anonymous
    Species: Bovine
    Assay Type: In Vitro
    Cell Line/Tissue: chondrocyte
    Verified Customer | Posted 03/26/2018
    I dissolved in DMSO
    Y-27632 dihydrochloride 1254
  • I prepare 20mg of the product in 5.88mL H2O and I use 1uL/mL of the stock in my cell culture after the cell splitting
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: human foreskin fibroblasts-derived-IPS
    Verified Customer | Posted 02/01/2018
    Keep the stock frozen at -20°C. I use it freshly added to the medium when splitting the cells.
    I used it in two human IPS cell lines, 5uM concentration the first 24 h after the splitting. The use of this reagent improves substantially the cell survival.
    Y-27632 dihydrochloride 1254
  • ROCK Inhibitor to improve cell survival
    Name: Hudson DE OLIVEIRA BEZERRA
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: H1 (W01)
    Verified Customer | Posted 01/05/2018
    My team uses this product to improve human embryonic stem cells survival in single cell passage methods in siRNA and CRISPR experiments. In general, we use the concentration of 10 uM. The products work very well, better than others available in the market.
    Y-27632 dihydrochloride 1254

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