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Submit ReviewY-27632 dihydrochloride is a selective ROCK inhibitor (Ki values are 0.14-0.22, 0.3, 25, 26 and > 250 μM for ROCK1 (p160 ROCK), ROCK2, PKA, PKC and MLCK respectively). Also inhibits PRK2 (IC50 = 600 nM). Increases survival rate of human embryonic stem (hES) cells and iPSC undergoing cryopreservation. Used in combination with CHIR 99021, RepSox, Forskolin, SP 600125, Go 6983 and VPA to reprogram fibroblasts to mature neurons. Also optimizes naive human pluripotent stem cell growth and viability following naive cell derivation from primed ESCs and iPSCs using naive human stem cell medium (NHSM). Used as component of growth media for urothelial organoids. Also used in a protocol to generate brain organoids from human iPSCs. Cell-permeable, orally bioavailable and active in vivo.
Y-27632 in solution and Y-27632 synthesized to cGMP guidelines also available.
For more information about how Y-27632 dihydrochloride may be used, see our protocols: Transdifferentiating Fibroblasts into Neurons, Converting Fibroblasts into Cardiomyocytes (9C Cocktail), Accelerated Induction of Cortical Neurons from hiPSCs, Human iPSC-derived Myoblasts.
Application of Y-27632 in MESP1+ Mesoderm. ROCK inhibition by Y-27632 (1 μM) promotes skeletal myogenic differentiation of MESP1+ mesoderm, as shown by immunostaining for Myosin Heavy Chain+ (Green) and Myogenin (Red), and counterstaining with DAPI (Catalog # 5748; Blue, Tocris). Data courtesy of Dr Sunny Chan and Dr Michael Kyba, University of Minnesota.
Y-27632 dihydrochloride is also offered as part of the Tocriscreen 2.0 Max, Tocriscreen Kinase Inhibitor Library and Tocriscreen Stem Cell Library. Find out more about compound libraries available from Tocris.
M. Wt | 320.26 |
Formula | C14H21N3O.2HCl |
Storage | Desiccate at RT |
Purity | ≥98% (HPLC) |
CAS Number | 129830-38-2 |
PubChem ID | 14801906 |
InChI Key | IDDDVXIUIXWAGJ-LJDSMOQUSA-N |
Smiles | N[C@H](C)[C@@]1([H])CC[C@H](C(NC2=CC=NC=C2)=O)CC1.Cl.Cl |
The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.
Tocris products are intended for laboratory research use only, unless stated otherwise.
Solvent | Max Conc. mg/mL | Max Conc. mM | |
---|---|---|---|
Solubility | |||
water | 32.03 | 100 | |
DMSO | 32.03 | 100 |
The following data is based on the product molecular weight 320.26. Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
Concentration / Solvent Volume / Mass | 1 mg | 5 mg | 10 mg |
---|---|---|---|
1 mM | 3.12 mL | 15.61 mL | 31.22 mL |
5 mM | 0.62 mL | 3.12 mL | 6.24 mL |
10 mM | 0.31 mL | 1.56 mL | 3.12 mL |
50 mM | 0.06 mL | 0.31 mL | 0.62 mL |
References are publications that support the biological activity of the product.
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Santos et al (2019) Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity. Nat.Commun. 10 4407 PMID: 31562298
Schafer (2023) An in vivo neuroimmune organoid model to study human microglia phenotypes. Cell 186 2111 PMID: 37172564
If you know of a relevant reference for Y-27632 dihydrochloride, please let us know.
Keywords: Y-27632 dihydrochloride, Y-27632 dihydrochloride supplier, Selective, p160, ROCK, inhibitors, inhibits, Rho-Kinase, Rho-Associated, Coiled-Coil, Kinases, ROK, stem, cells, organoids, induced, pluripotent, ROCKI, ROCK1, ROCKII, ROCK2, reprogramming, cryopreservation, differentiation, cell, freezing, media, 2HCl, y27632, Y-27632, inhibitor, Rho-kinases, Stem, Cell, Proliferation, Organoids, Reprogramming, Neural, Cells, 1254, Tocris Bioscience
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ROCKi has consistently performed in iPSC cell culture work. Well packaged vials are resuspended and aliquoted for -20 storage. Cells passaged or fed with ROCKi media have shown good viability. This is a regular purchase for the labs at a reasonable price.Images @10x magnification show limited differentiation of iPSCs after 3 days in culture.
We add this product at a concentration of 10 uM to promote cell survival in in vitro iPSC cultures.
Easy to manage and works as expected
To generate human intestinal organoids
I used this product on freshly split and plate human iPSC cells to help cell survival and adherence. I used the product at 10 mM concentration which I diluted in 1:1000 in culture media and left the cells at 37C, 5% CO2 for 24 hours before replacing with fresh culture media.
For hPSC culture, final concentration 10mM. Reconstitute 10 mg of Y27632 in 3.0792 ml of PBS. Make 100µl aliquots and keep them at −20 °C for up to 6 months.
HPLC
ROCK inhibitor Y-27632 (10 mM)
Great for passaging huiPSCs with dissociation into single cells.
Used during single cell passage for CRISPR clone isolation, works well and no difference found between multiple batches. Cells show characteristic 'spiky' morphology upon addition to the medium.
I use at 1:1000 and store at -20C. I remove the compound from medium 24 hours after plating.
For iPSC culture, final concentration 10mM. Reconstitute 10 mg of Y27632 in 3.0792 ml of PBS.
Make 100µl aliquots and keep them at −20 °C for up to 6 months.
I dissolved in DMSO
I used it in two human IPS cell lines, 5uM concentration the first 24 h after the splitting. The use of this reagent improves substantially the cell survival.
Keep the stock frozen at -20°C. I use it freshly added to the medium when splitting the cells.
My team uses this product to improve human embryonic stem cells survival in single cell passage methods in siRNA and CRISPR experiments. In general, we use the concentration of 10 uM. The products work very well, better than others available in the market.
The following protocols feature additional information for the use of Y-27632 dihydrochloride (Cat. No. 1254).
Tocris offers the following scientific literature in this area to showcase our products. We invite you to request* your copy today!
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Written by Kirsty E. Clarke, Victoria B. Christie, Andy Whiting and Stefan A. Przyborski, this review provides an overview of the use of small molecules in the control of stem cell growth and differentiation. Key signaling pathways are highlighted, and the regulation of ES cell self-renewal and somatic cell reprogramming is discussed. Compounds available from Tocris are listed.
Stem cells have potential as a source of cells and tissues for research and treatment of disease. This poster summarizes some key protocols demonstrating the use of small molecules across the stem cell workflow, from reprogramming, through self-renewal, storage and differentiation to verification. Advantages of using small molecules are also highlighted.