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Y-27632 dihydrochloride

Cat. No. M1817

All AbMole products are for research use only, cannot be used for human consumption.

Y-27632 dihydrochloride Structure
Synonym:

Y27632; Y-27632 2HCl

Size Price Availability Quantity
Free Sample (0.5-1 mg)  USD 0 In stock
10mM*1mL in DMSO USD 45 In stock
2mg USD 35 In stock
5mg USD 47 In stock
10mg USD 67 In stock
25mg USD 100 In stock
50mg USD 170 In stock
100mg USD 255 In stock
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Quality Control & Documentation
Biological Activity

Y-27632 is a potent, selective inhibitor of Rho-associated protein kinases (ROCK) with IC50 values of 140-220 nM for ROCK1 and ROCK2. Y-27632 inhibited the kinase activity of both ROCK-I and ROCK-II in vitro, and this inhibition was reversed by ATP in a competitive manner. The ROCK family of Rho-associated serine-threonine protein kinases is known to play an important role in Rho-mediated cell adhesion and smooth muscle contraction. It also inhibits PRK2 with an IC50 value of 600 nM. One-hour treatment with 10 µM Y-27632 blocks apoptosis of dissociated cultured human embryonic stem cells, increasing cloning efficiency by 25% and sustaining survival up to 30 passages. Y-27632 was found to increases human embryonic stem cell (hESC) survival. In addition, Y-27632 inhibited LPA-induced entosis.

Customer Product Validations & Biological Datas
Application Cytoskeleton and nucleus staining in the absence or presence of ROCK inhibitor (Y-27632)
Results The attached cells on the three investigated surfaces in the presence or absence (control) of Y-27632 after 4 h and 24 h were summarized in Fig. 7
Application Cytoskeleton and nucleus staining in the absence or presence of ROCK inhibitor (Y-27632)
Results After inhibition of ROCK by Y-27632, an expected formation of finger-like filopodia was seen for the cells of all groups after 4 h, as shown by white arrows in Fig. 8b, f and j.
Application Cell culture.
Results
Application MTT assay
Results Absence of the ROCK inhibitor (Y-27632) will greatly decrease the efficiency of organoid outgrowth.
Application Organoid culture
Results A few cells presented as an aggregate of two cells or three cells. At day 7, LNCaP cells and C4-2B cells started to present aggregates of multiple cells. At day 14, organoids were formed as aggregates of LNCaP cells and C4-2B cells
Application PARPi assay
Results Inhibition of Rho-associated coiled-coil containing protein kinase (ROCK) has been shown to allow long-term proliferation of tumor epithelial cells in vitro and addition of the specific ROCK inhibitor Y-27632 indeed improved culture conditions.
Application FACS plots
Results FACS plots depicting gating strategies for sorting single cells from a human liver organoid culture
Application Patient-derived organoid culture
Results mCRPC patient-derived organoids are sensitive to olaparib. Organoids from patients with homozygous loss of CHD1 (V5272 and V5372) show increased sensitivity to PARP inhibition.
Application i.e.
Results Administration of tamoxifen to these mice induces specific activation of the Cre enzyme within the epithelial cells of the small and large intestine, with consequential deletion of both Apc alleles, expression of an oncogenic form of Kras (KrasG12D) and deletion of one of the two Trp53 alleles
Application Transwell
Results Y27632 did not affect the scattering and invasion of MDA-MB-231 cultured without CAFs, but significantly reduced the scattering and invasion induced by CAFs, suggesting that CAFs promote the invasion of MDA-MB-231 cells via ROCK1/2.
Application immunoblotting
Results Interestingly, PAI-1 expression increased in cells treated with Y27632, suggesting the existence of a regulatory loop between PAI-1 and ROCK
Application intraperitoneally
Results After 18 days of treatment, we did not observe any effect of PQ401, Y27632 or PQ401 combined with Y27632 on primary tumor growth, suggesting that the inhibition of ROCK or IGF-1 receptor did not affect cancer cell proliferation.
Application intraperitoneally
Results Similar to our in vitro findings, Y27632 significantly increased the expression of PAI-1, confirming the existence of a regulatory mechanism between ROCK and PAI-1 in vivo
Application cell culture
Results Lastly, IFA of sporozoiteinfected organoids using Crypt-a-Glo showed newly formed oocysts at six days post infection
Application Organoid Isolation and Culturing
Results The frequency of cyst formation from single intestinal stem cells after three days (colony formation efficiency) was equally high in the fibrin gel supplemented with 10% v/v BME as in pure BME.
Application general culture
Results Illustration of organoid generation from tumor, to plating in Matrigel, to organoid growth
Protocol (for reference only)
Cell Experiment
Cell lines cyES cells
Preparation method For 5-bromo-2′-deoxyuridine (BrdU) incorporation assays, dissociated cyES cells were seeded at a concentration of 500 cells/cm2 onto fresh feeder layers in the presence of the ROCK inhibitors Y-27632 (Wako Pure Chemical Industries, Tokyo, Japan) or Fasudil (Tocris Bioscience, MO, USA) during the first 24 h of culture. The BrdU incorporation assay was performed using colorimetric BrdU Cell proliferation Kit (Roche Diagnostics) according to the manufacturer's instructions. BrdU incorporations were also observed by immunocytochemical (ICC) staining with BrdU Labeling and Detection Kit I (Roche Diagnostics).
Concentrations 0, 0.1, 0.5, 5, 10, 20µM
Incubation time 24 h
Animal Experiment
Animal models EAC modeling
Formulation dissolved in 0.9% NaCl
Dosages 0.1, 1, 10 mg/kg/day for 14 days
Administration i.p.
Chemical Information
Molecular Weight 320.26
Formula C14H21N3O.2HCl
CAS Number 129830-38-2
Solubility (25°C) Water ≥ 60 mg/mL
DMSO ≥ 35 mg/mL
Storage Powder          -20°C   3 years ;  4°C   2 years
In solvent       -80°C   6 months ;  -20°C   1 month
References

[1] Ishizaki T, et al. Mol Pharmacol. Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases.

[2] Honjo M, et al. Invest Ophthalmol Vis Sci. Effects of rho-associated protein kinase inhibitor Y-27632 on intraocular pressure and outflow facility.

[3] Li X, et al. Stem Cells Dev. The ROCK inhibitor Y-27632 enhances the survival rate of human embryonic stem cells following cryopreservation.

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Keywords: Y-27632 dihydrochloride, Y27632; Y-27632 2HCl supplier, ROCK, inhibitors, activators

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