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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 70  USD70 In stock
2mg USD 40  USD40 In stock
5mg USD 55  USD55 In stock
10mg USD 90  USD90 In stock
50mg USD 240  USD240 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.

Product Citations
Customer Product Validations & Biological Datas
Source Cancer Discov (2018 Sep). Figure 1. Y27632 (AbMole BioScience)
Method general culture
Cell Lines Pleural effusion fluid
Concentrations 10 uM
Incubation Time
Results Illustration of organoid generation from tumor, to plating in Matrigel, to organoid growth
Source Advanced materials (2018 Oct). Figure 1. Y-27632 (Abmole Bioscience)
Method Organoid Isolation and Culturing
Cell Lines Mouse small intestinal organoids
Concentrations 10 × 10−6 M
Incubation Time 20-45 min
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.
Source Nature Microbiology (2018). Figure 3. Y27632 (AbMole BioScience)
Method cell culture
Cell Lines Lung tissue
Concentrations 10 uM
Incubation Time 2 weeks
Results Lastly, IFA of sporozoiteinfected organoids using Crypt-a-Glo showed newly formed oocysts at six days post infection
Source Oncotarget (2017). Figure 6. Y27632 (Abmole Bioscience)
Method intraperitoneally
Cell Lines SCID mice
Concentrations 25 mg/kg
Incubation Time 18 days
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
Source Oncotarget (2017). Figure 5. Y27632 (Abmole Bioscience)
Method intraperitoneally
Cell Lines SCID mice
Concentrations 25 mg/kg
Incubation Time 18 days
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.
Source Oncotarget (2017). Figure 4. Y27632 (Abmole Bioscience)
Method immunoblotting
Cell Lines MDA-MB-231 cell
Concentrations 10 μM
Incubation Time 72 h
Results Interestingly, PAI-1 expression increased in cells treated with Y27632, suggesting the existence of a regulatory loop between PAI-1 and ROCK
Source Oncotarget (2017). Figure 2. Y27632 (Abmole Bioscience)
Method Transwell
Cell Lines MDA-MB-231 cells
Concentrations 10 μM
Incubation Time 72 h
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.
Source Nat Protoc (2018). Figure 1. Y-27632 (Abmole Bioscience)
Method i.e.
Cell Lines NSG (NOD.Cg-Prkd cscidIl 2rgtm1Wjl/SzJ) mice
Concentrations 10 μM
Incubation Time 5–10 min
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
Source University of California (2017). Figure 7. Y-27632 (Abmole Bioscience)
Method Patient-derived organoid culture
Cell Lines biopsy tissue
Concentrations 10 μM
Incubation Time 30 min
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.
Source Nat Protoc (2018). Figure 4. Y-27632 dihydrochloride (Abmole Bioscience, USA)
Method FACS plots
Cell Lines Human liver single-cell
Concentrations 10 μM
Incubation Time 7 d
Results FACS plots depicting gating strategies for sorting single cells from a human liver organoid culture
Source Cell (2017). Figure 1. Y-27632 (Abmole Bioscience)
Method PARPi assay
Cell Lines BC organoid lines
Concentrations < 100 μm
Incubation Time 48 h
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.
Source Am J Clin Exp Urol (2017) . Figure 1. Y-27632 (Abmole Bioscience, USA)
Method Organoid culture
Cell Lines LNCaP and C4-2B cells
Concentrations 10 μM
Incubation Time 48-72 h
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
Source Nature Protocols (2016) . Figure 3. Y-27632 (Abmole Bioscience)
Method MTT assay
Cell Lines mouse and human luminal cell
Concentrations 10 μM
Incubation Time 2–3 d
Results Absence of the ROCK inhibitor (Y-27632) will greatly decrease the efficiency of organoid outgrowth.
Source Nature (2016). Figure 1. Y-27632 (Abmole Bioscience Inc)
Method Cell culture.
Cell Lines single cells
Concentrations 10 μM
Incubation Time
Results
Source Materials Science and Engineering (2017). Figure 8. Y-27632 (Abmole Bioscience Inc, USA)
Method Cytoskeleton and nucleus staining in the absence or presence of ROCK inhibitor (Y-27632)
Cell Lines SaOS-2 cells
Concentrations 10 μM
Incubation Time 4 h and 24 h
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.
Source Materials Science and Engineering (2017). Figure 7. Y-27632 (Abmole Bioscience Inc, USA)
Method Cytoskeleton and nucleus staining in the absence or presence of ROCK inhibitor (Y-27632)
Cell Lines SaOS-2 cells
Concentrations 10 μM
Incubation Time 4 h and 24 h
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
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] Li X, et al. Stem Cells Dev. The ROCK inhibitor Y-27632 enhances the survival rate of human embryonic stem cells following cryopreservation.

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

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

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