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 426 publications

Ancestral allele of DNA polymerase gamma modifies antiviral tolerance [ Nature, 2024, 10.1038/s41586-024-07260-z]	PubMed: 38570685
Ancestral allele of DNA polymerase gamma modifies antiviral tolerance [ Nature, 2024, 628(8009):844-853]	PubMed: 38570685
Vitamin A resolves lineage plasticity to orchestrate stem cell lineage choices [ Science, 2024, 383(6687):eadi7342]	PubMed: 38452090
Osteopontin drives neuroinflammation and cell loss in MAPT-N279K frontotemporal dementia patient neurons [ Cell Stem Cell, 2024, S1934-5909(24)00094-8]	PubMed: 38626772
Simultaneous proteome localization and turnover analysis reveals spatiotemporal features of protein homeostasis disruptions [ Nat Commun, 2024, 15(1):2207]	PubMed: 38467653
Bioelectric stimulation controls tissue shape and size [ Nat Commun, 2024, 15(1):2938]	PubMed: 38580690
Octyl itaconate enhances VSVΔ51 oncolytic virotherapy by multitarget inhibition of antiviral and inflammatory pathways [ Nat Commun, 2024, 15(1):4096]	PubMed: 38750019
Zebrafish Avatar-test forecasts clinical response to chemotherapy in patients with colorectal cancer [ Nat Commun, 2024, 15(1):4771]	PubMed: 38839755
Octyl itaconate enhances VSVΔ51 oncolytic virotherapy by multitarget inhibition of antiviral and inflammatory pathways [ Nat Commun, 2024, 15(1):4096]	PubMed: 38750019
High-speed optical imaging with sCMOS pixel reassignment [ Nat Commun, 2024, 15(1):4598]	PubMed: 38816394

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