Y-27632

Catalog No.S6390 Batch:S639001

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
* <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.)

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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.^[2]

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.^[3]

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

Selleck's Y-27632 has been cited by 522 publications

Highly efficient construction of monkey blastoid capsules from aged somatic cells [ Nat Commun, 2025, 16(1):1130]	PubMed: 39875393
Frictiotaxis underlies focal adhesion-independent durotaxis [ Nat Commun, 2025, 16(1):3811]	PubMed: 40268931
BRD8 Guards the Pluripotent State by Sensing and Maintaining Histone Acetylation [ Adv Sci (Weinh), 2025, 12(5):e2409160]	PubMed: 39656858
Combining molecular characteristics and therapeutic analysis of PDOs predict clinical responses and guide PDAC personalized treatment [ J Exp Clin Cancer Res, 2025, 44(1):72]	PubMed: 40001264
Sulforaphane protects developing neural networks from VPA-induced synaptic alterations [ Mol Psychiatry, 2025, 10.1038/s41380-025-02967-5]	PubMed: 40175519
Functional genomics pipeline identifies CRL4 inhibition for the treatment of ovarian cancer [ Clin Transl Med, 2025, 15(2):e70078]	PubMed: 39856363
The modulation of calcium and chloride channels induces cardiomyocytes from human pluripotent stem cells [ Int J Biol Sci, 2025, 21(1):95-108]	PubMed: 39744425
Identification and validation of a T cell receptor targeting KRAS G12V in HLA-A*11:01 pancreatic cancer patients [ JCI Insight, 2025, 10(2)e181873]	PubMed: 39846249
Comparison of characteristics and immune responses between paired human nasal and bronchial epithelial organoids [ Cell Biosci, 2025, 15(1):18]	PubMed: 39920853
Super-enhancer-hijacking RBBP7 potentiates metastasis and stemness of breast cancer via recruiting NuRD complex subunit LSD1 [ J Transl Med, 2025, 23(1):266]	PubMed: 40038738

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