Y-27632

Catalog No.S6390 Batch:S639002

Technical Data

Formula

C₁₄H₂₁N₃O

Molecular Weight

247.34

CAS No.

146986-50-7

Solubility (25°C)*

In vitro

DMSO

49 mg/mL (198.1 mM)

Ethanol

49 mg/mL (198.1 mM)

Water

Insoluble

In vivo (Add solvents to the product individually and in order)

Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	2.500mg/ml (10.11mM)	Taking the 1 mL working solution as an example, add 50 μL of 50 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

Y-27632 is a selective ROCK1 and ROCK2 inhibitor with a Ki of 140 nM and 300nM in a cell-free assay, exhibits >200-fold selectivity over other kinases, including PKC, cAMP-dependent protein kinase, MLCK and PAK.

Targets

ROCK1 ^[1] (Cell-free assay)	ROCK2 ^[1] (Cell-free assay)
140 nM(Ki)	300 nM(Ki)

In vitro

Y-27632 primes human induced pluripotent stem cells (hIPSCs) to selectively differentiate towards mesendodermal lineage via epithelial-mesenchymal transition-like modulation.

In vivo

Y27632, a specific inhibitor of ROCK, supresses ROCK as well as altered pathways and leads to a significant enhancement in corneal nerve regeneration.

Protocol (from reference)

Cell Assay:^{^[2]}	Cell lines Human iPSC line nciPS02, RC01001-B, Female; Pluripotent stem cells Concentrations 20 µM Incubation Time -- Method The hiPSC line is cultured on six-well plates coated with 1% Matrigel. Pluripotent stem cells are maintained in mTeSR1 (Stem Cell) medium supplemented with 100 µg/mL Normocin coated with 1% Matrigel. For differentiation, hiPSC colonies are detached from the culture dish using Accutase Cell Dissociation Reagent. Then, cells are collected as a single-cell suspension in Essential 8 Flex medium containing 20 µm Y27632. Cells are distributed at a density of 3500 cells in 100 µL of medium per well into 96-well U-bottom plates. After 24 h of incubation at 37 ℃, fresh E8 medium without Y27632 is added to a total volume of 200 µL per well, and the plates are incubated at 37℃ continuously. Next, cell aggregates are cultured in E6-based differentiation medium containing 2% Matrigel, 10 µm SB431542, 4 ng mL 1 basic FGF, and 2.5 ng mL 1 BMP4. On day 3 of differentiation, 200 ng mL 1 LDN-193189 and 50 g mL 1 FGF are added to induce cranial neural crest cell formation. On day 12, all cell aggregates are transferred to 24-well low-attachment plates in 500 µL of organoid maturation mediumcontaining 1% Matrigel to induce self-assembly of the epidermis. Half of the medium is replaced after 16 d after differentiation. On day 17, the organoids are collected for the subsequent experiments.
Animal Study:^{^[3]}	Animal Models C57BL/6J mice Dosages 5 nM Administration eye drops

Cell Assay:^{^[2]}

Cell lines
Human iPSC line nciPS02, RC01001-B, Female; Pluripotent stem cells
Concentrations
20 µM
Incubation Time
--
Method
The hiPSC line is cultured on six-well plates coated with 1% Matrigel. Pluripotent stem cells are maintained in mTeSR1 (Stem Cell) medium supplemented with 100 µg/mL Normocin coated with 1% Matrigel. For differentiation, hiPSC colonies are detached from the culture dish using Accutase Cell Dissociation Reagent. Then, cells are collected as a single-cell suspension in Essential 8 Flex medium containing 20 µm Y27632. Cells are distributed at a density of 3500 cells in 100 µL of medium per well into 96-well U-bottom plates. After 24 h of incubation at 37 ℃, fresh E8 medium without Y27632 is added to a total volume of 200 µL per well, and the plates are incubated at 37℃ continuously. Next, cell aggregates are cultured in E6-based differentiation medium containing 2% Matrigel, 10 µm SB431542, 4 ng mL 1 basic FGF, and 2.5 ng mL 1 BMP4. On day 3 of differentiation, 200 ng mL 1 LDN-193189 and 50 g mL 1 FGF are added to induce cranial neural crest cell formation. On day 12, all cell aggregates are transferred to 24-well low-attachment plates in 500 µL of organoid maturation mediumcontaining 1% Matrigel to induce self-assembly of the epidermis. Half of the medium is replaced after 16 d after differentiation. On day 17, the organoids are collected for the subsequent experiments.

Animal Study:^{^[3]}

Animal Models
C57BL/6J mice
Dosages
5 nM
Administration
eye drops

References

https://pubmed.ncbi.nlm.nih.gov/9353125/
https://pubmed.ncbi.nlm.nih.gov/35315234/
https://pubmed.ncbi.nlm.nih.gov/34419637/

Selleck's Y-27632 Has Been Cited by 582 Publications

MeCP2 gene dosage-dependent neurodevelopmentally restricted defects arise by aberrant activation of cell fate-determining bivalent genes [ Nat Commun, 2026, 17(1)3225]	PubMed: 42026056
zAvatar-test-A functional precision model to personalize ovarian cancer treatments: Results from a co-clinical study [ Cell Rep Med, 2026, 7(1):102530]	PubMed: 41475339
BMP4 initiates and patterns ventral-caudal structures in zebrafish and human pluripotent stem cell aggregates [ EMBO J, 2026, 45(1):210-242]	PubMed: 41286028
CRISPR-MiX: A pooled single-stranded donor strategy to enhance HDR efficiency in human iPSCs [ Mol Ther Nucleic Acids, 2026, 37(1):102820]	PubMed: 41674724
Pharmacological strategies to enhance the response of hepatoblastoma to chemotherapy through MDR1 inhibition [ Acta Pharmacol Sin, 2026, 10.1038/s41401-026-01791-z]	PubMed: 42026237
NAD+ augmentation by nicotinamide riboside engages SLIT2/ROBO1 signaling to attenuate Th17 inflammation in psoriasis [ JCI Insight, 2026, e203826]	PubMed: 42048163
Early hypoxia treatment and oxygen-concentration alteration promote the retinal progenitor proliferation and ganglion cell maturation in human retinal organoid development [ Stem Cell Res Ther, 2026, 17(1)109]	PubMed: 41664121
Head and Neck Squamous Cell Carcinoma Patient-Derived Cancer-Associated Fibroblasts Undergo Senescence While Retaining Protumorigenic Properties After Ex Vivo Irradiation [ Int J Radiat Oncol Biol Phys, 2026, S0360-3016(26)00395-0]	PubMed: 41698472
Spatially Resolved Proteomic Mapping in Skin Organoid for Hair Follicle Development [ Mol Cell Proteomics, 2026, 25(1):101482]	PubMed: 41380997
ADAMDEC1-driven CCL2-CCR2-PD-1 axis in esophageal squamous cell carcinoma: a dual threat of O-GlcNAcylation and m6A methylation in neoadjuvant therapy resistance and immune evasion [ Cell Oncol (Dordr), 2026, 49(2)68]	PubMed: 41944985

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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