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)

Clear solution

5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O

2.5mg/ml

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.^[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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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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