Y-27632

N.º de catálogoS6390 Lote:S639001

Imprimir

Datos técnicos

Fórmula

C₁₄H₂₁N₃O

Peso molecular

247.34

Número CAS

146986-50-7

Solubilidad (25°C)*

In vitro

DMSO

49 mg/mL (198.1 mM)

Ethanol

49 mg/mL (198.1 mM)

Water

Insoluble

In vivo (Agregue los solventes al producto individualmente y en orden.)

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.

* <1 mg/ml significa ligeramente soluble o insoluble.
* Tenga en cuenta que Selleck prueba la solubilidad de todos los compuestos internamente, y la solubilidad real puede diferir ligeramente de los valores publicados. Esto es normal y se debe a ligeras variaciones entre lotes.
* Envío a temperatura ambiente (Las pruebas de estabilidad demuestran que este producto se puede enviar sin medidas de refrigeración.)

Preparación de soluciones madre

Actividad biológica

Descripción

Y-27632 es un inhibidor selectivo de ROCK1 y ROCK2 con una Ki de 140 nM y 300 nM en un ensayo sin células, que exhibe una selectividad >200 veces mayor sobre otras quinasas, incluyendo PKC, proteína quinasa dependiente de cAMP, MLCK y PAK.

Objetivos

ROCK1 (Cell-free assay)	ROCK2 (Cell-free assay)
140 nM(Ki)	300 nM(Ki)

In vitro

Y-27632 prepara las células madre pluripotentes inducidas humanas (hIPSCs) para diferenciarse selectivamente hacia el linaje mesendodérmico a través de una modulación similar a la transición epitelio-mesenquimatosa.

In vivo

Y27632, un inhibidor específico de ROCK, suprime ROCK, así como las vías alteradas, y conduce a una mejora significativa en la regeneración del nervio corneal.

Protocolo (de referencia)

Ensayo celular:^{^[2]}	Líneas celulares Human iPSC line nciPS02, RC01001-B, Female; Pluripotent stem cells Concentraciones 20 µM Tiempo de incubación -- Método 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.
Estudio en animales:^{^[3]}	Modelos animales C57BL/6J mice Dosificaciones 5 nM Administración eye drops

Ensayo celular:^{^[2]}

Líneas celulares
Human iPSC line nciPS02, RC01001-B, Female; Pluripotent stem cells
Concentraciones
20 µM
Tiempo de incubación
--
Método
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.

Estudio en animales:^{^[3]}

Modelos animales
C57BL/6J mice
Dosificaciones
5 nM
Administración
eye drops

Referencias

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

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