Y-27632 2HCl

For research use only.

Catalog No.S1049

429 publications

Y-27632 2HCl Chemical Structure

Molecular Weight(MW): 320.26

Y-27632 2HCl is a selective ROCK1 (p160ROCK) inhibitor with Ki of 140 nM in a cell-free assay, exhibits >200-fold selectivity over other kinases, including PKC, cAMP-dependent protein kinase, MLCK and PAK.

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Selleck's Y-27632 2HCl has been cited by 429 publications

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Choose Selective ROCK Inhibitors

Biological Activity

Description Y-27632 2HCl is a selective ROCK1 (p160ROCK) inhibitor with Ki of 140 nM in a cell-free assay, exhibits >200-fold selectivity over other kinases, including PKC, cAMP-dependent protein kinase, MLCK and PAK.
Targets
ROCK1 (p160ROCK) [1]
(Cell-free assay)
ROCK2 [6]
(Cell-free assay)
140 nM(Ki) 300 nM(Ki)
In vitro

Y-27632 2HCl inhibits ROCK-II while displaying little activity against PKC, cAMP-dependent protein kinase and myosin light-chain kinase (MLCK) with Ki of 26 μM, 25 μM and > 250 μM, respectively, as well as PKA activated by another Rho-family GTPase member, Cdc42. Y-27632 2HCl inhibits smooth-muscle contraction induces by various agonists including phenylephrine, histamine, acetylcholine, serotonin, endothelin, and thromboxane with IC50 of 0.3-1 μM, by selectively inhibiting Ca2+ sensitization. Y-27632 2HCl suppresses Rho-induced, p160ROCK-mediated formation of stress fibres in cultured cells. [1] Y-27632 2HCl treatment blocks both Rho-mediated activation of actomyosin and LPA-stimulated invasive activity of MM1 cells in a concentration-dependent manner. [2] Y-27632 2HCl treatment is not only sufficient to initiate formation of exuberant axonal processes but also facilitates axonal maturation during the very early stages of axonogenesis, while largely sparing axon elongation. [3] In human embryonic stem (hES) cells, Y-27632 2HCl treatment at 10 μM markedly diminishes dissociation-induced apoptosis even in serum-free suspension (SFEB) culture, increases cloning efficiency (from ~1% to ~27%), facilitates subcloning after gene transfer, and enables SFEB-cultured hES cells to survive and differentiate into Bf1+ cortical and basal telencephalic progenitors. [4]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Swiss 3T3 cells M3fOdGZ2dmO2aX;uJGF{e2G7 MmL3NVAh|ryP NGrJV5ozKGh? NHfBVIFFVVOR MYrJcohq[mm2czD0bIUh[XO|ZX3icJkhd2ZibXnjdo91fWK3bHXzJIFv\CCrboTldo1m\GmjdHWg[olt[W2nboTzJJRwKG[xcn2g[Zh1\W6mZXSgdJJw[2W|c3Xz M3TVeFk3PDd4NUS=
N1E-115 NFPHd25HfW6ldHnvckBCe3OjeR?= NUjEfoFiOTBizszN MWOyJIg> MmnmSG1UVw>? Ml7sTY5pcWKrdIOgeIhmKGG|c3XtZox6KG:oIH3pZ5JwfHWkdXzld{BidmRiaX70[ZJu\WSrYYTlJIZqdGGvZX70d{B1dyCob4LtJIV5fGWwZHXkJJBzd2Onc4Pldy=> M3fYfVk3PDd4NUS=
HeLa MkXESpVv[3Srb36gRZN{[Xl? M3LQTVExKM7:TR?= MoSwN|AhdWmw Mn\ETY5pcWKrdIOgeIhmKG[xcn3heIlwdiCxZjDzeJJme3NiZnni[ZJ{KGGwZDD0bIUh[XO|ZX3icJkhd2ZidnnuZ5VtcW5vY3;ueIFqdmmwZzDmc4NidCCjZHjld4lwdnN? MVy5OlY5ODd{
CCL39 M1\5fWZ2dmO2aX;uJGF{e2G7 NHXObpY{OCEQvF2= NG\XW|I{OCCvaX6= M1Wx[mNwdXCuZYTlcJkh[WKxbHnzbIV{KGGldHn2ZZRqd25ib3[gUoEuUCCneHPoZY5o\XJiTljFNUBjgSCrboTl[5JqdnN? NXrXc3U5QTZ7M{O4Ni=>
Mesothelial cells from rat mesentery MVrJcpZie2m4ZTDBd5NigQ>? NFXwUGE{OCEQvF2= NEf0S|MzOCCq MmWwRoxw[2u|IHnueoF{cX[nIHHjeIl3cXS7 MmrYPVk{ODh5Mh?=
NIH3T3 MWrHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MmXUNVAh|ryP MorKNVgh\A>? MWfEc4V{KG6xdDDpcohq[mm2IHPlcIwh\3Kxd4To NWexVIZJOTByMkGzPFY>
Dbl-d M4DFbmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NHvWRogyOCEQvF2= M3\qZ|E5KGR? M3zFOnN1em:wZ3z5JIlvcGmkaYTzJINmdGxiZ4Lve5Rp MYOxNFAzOTN6Nh?=
Dbl-e MmSyS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MYWxNEDPxE1? MYmxPEBl MnXrUY9l\XKjdHXsfUBqdmirYnn0d{Bk\WyuIHfyc5d1cA>? NVT5PItNOTByMkGzPFY>
mNET1-d MUDHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MUexNEDPxE1? NVm2bWNnOThiZB?= MmjNV5Rzd26pbImgbY5pcWKrdIOgZ4VtdCCpcn;3eIg> MVSxNFAzOTN6Nh?=
mNET1-e MXjHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? Mn\ENVAh|ryP NYi1eYg1OThiZB?= MWnTeJJwdmeueTDpcohq[mm2czDj[YxtKGe{b4f0bC=> NHzvblEyODB{MUO4Oi=>
Ras-2 M4nFTGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M37QT|ExKM7:TR?= NU[3eW83OThiZB?= NV3YOpFQW3S{b37ncJkhcW6qaXLpeJMh[2WubDDndo94fGh? M4DUZlExODJzM{i2
Ras-4 MXnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MXixNEDPxE1? MkTTNVgh\A>? M{nv[XN1em:wZ3z5JIlvcGmkaYTzJINmdGxiZ4Lve5Rp M4HD[VExODJzM{i2
Src-1 MkW0S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M3XRNFExKM7:TR?= MojFNVgh\A>? NXKwdJN1TG:nczDuc5QhcW6qaXLpeEBk\WyuIHfyc5d1cA>? MkjLNVAxOjF|OE[=
Src-4 NXjadpVKT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= M3znblExKM7:TR?= NEDsNGQyQCCm NIT1e5RFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp NVr2W5JIOTByMkGzPFY>
NIH3T3 MXHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NXSxRoR5OTBizszN MXqxPEBl NGPhcIdFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp MWexNFAzOTN6Nh?=
Src-1 MYHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NYXiW2tDOTBizszN Mk\PNVgh\A>? MkjESI9meyCwb4SgbY5pcWKrdDDj[YxtKGe{b4f0bC=> M2O3XFExODJzM{i2
Src-2 MWnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MYmxNEDPxE1? NFq3NYgyQCCm MYHEc4V{KG6xdDDpcohq[mm2IHPlcIwh\3Kxd4To MmTtNVAxOjF|OE[=
SW620 NFrDTGRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M2TIdVExKM7:TR?= NW\LVGJEOThiZB?= NGjKSFVFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp M2nSUVExODJzM{i2
HCT15 MULHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M2raZVExKM7:TR?= MojuNVgh\A>? NGHiXVFFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp NE\ifYsyODB{MUO4Oi=>
HCT116 NWXwV3QxT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NF72V24yOCEQvF2= MW[xPEBl MnTHV5Rzd26pbImgbY5pcWKrdIOgZ4VtdCCpcn;3eIg> NWXDZ|d{OTByMkGzPFY>
LS174T NFS3ZYhIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MlLsNVAh|ryP NULWOXo5OThiZB?= Mm\pUY9l\XKjdHXsfUBqdmirYnn0d{Bk\WyuIHfyc5d1cA>? MXmxNFAzOTN6Nh?=
Neonatal rat ventricular myocytes Mk\VSpVv[3Srb36gRZN{[Xl? MWWxNEDPxE1? NH7TWYk1QCCq NFqxNG5KdmirYnn0d{BGXC1zLXnu[JVk\WRiaX7jdoVie2W|IHnuJJBzd3SnaX6gd5lvfGinc3nzMEBk\WyuIIPpfoUh[W6mIH35c4Zq[nKrbHzhdkBwemejbnn6ZZRqd25? NITxcWkyODN6Nk[xNy=>
Stellate Cell NGLsVIpHfW6ldHnvckBCe3OjeR?= MYiyOUDPxE1? MX2xOUBucW5? MnrCTY5pcWKrdIOg[o9zdWG2aX;uJI9nKEZvYXP0bY4he3S{ZYPzJIZq[mW{czDhcoQheGixc4Doc5J6dGG2aX;uJI9nKG27b4PpckBtcWeqdDDjbIFqdg>? Ml\NNVA3ODB2OU[=
Rat Vascular Smooth Muscle Cells MWDGeY5kfGmxbjDBd5NigQ>? MUKxNEDPxE1? MXiyJIg> NH7lTVRKdmirYnn0d{Bidmerb4TlcpNqdiCLST3pcoR2[2WmIHj5dIVzfHKxcHj5 NWKyRoVzOTB4NEKzNVc>
PC3 MkK5SpVv[3Srb36gRZN{[Xl? NVLPcoRJOjVizszN NUKx[mZLOSCq MnnaTY5lfWOnczDtc5JxcG:ub3fpZ4FtKGOqYX7n[ZM> NI\jW|EyODd{MES3NS=>
PC3 M4DQUG1q\3KjdHnvckBCe3OjeR?= NVvCNoNIOjVizszN MnnQNUBp MUDJcohq[mm2czD0bIUhSk2IQj3DUUBidmRidHjlJGVITi2|dHnteYxifGWmIH3p[5JifGmxbh?= MmnhNVA4OjB2N{G=
PC3 MYLHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NF;BcYwzPSEQvF2= MlKwNVchcA>? MkDJSI9meyCwb4SgbY5pcWKrdDDj[YxtKGe{b4f0bC=> Ml\iNVA4OjB2N{G=
LNCaP NHfBcIdIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NHTqPZozPSEQvF2= NV3ZOGRxOTdiaB?= MULEc4V{KG6xdDDpcohq[mm2IHPlcIwh\3Kxd4To NUW1RnkzOTB5MkC0O|E>
Rat hepatic stellate cells MUfGeY5kfGmxbjDBd5NigQ>? Mk\wN|Ah|ryP MVy0PEBp MXPEbY1qdmm|aHXzJJRp\SCyaH;zdIhwenmuYYTpc44hd2ZiRYLrNkwh[W6mIHTlZ5Jm[XOnczDu[ZchTE6DIIP5cpRp\XOrcx?= MUSxNFg1PTZ4Mx?=
Pancreatic acinar cells MmG1SpVv[3Srb36gRZN{[Xl? NITHW4wyOMLizszN NEH4WWs4OCCvaX6= NGizd2dRd3SnboTpZZRmeyCFQ1utd5RqdXWuYYTl[EBx[W6lcnXheIlkKGWweont[UB{\WO{ZYTpc44> NFP1T3EyOjd2NUC4NC=>
C2C12 NFe2e2FHfW6ldHnvckBCe3OjeR?= MVOxNOKh|ryP NImwPW83KGh? NUPjOXVYWHKndnXueJMhfGinIIPldolv\SCyaH;zdIhwenmuYYTpc44hd2ZiSWLTMVEhcW6mdXPl[EBjgSCrboP1cIlvKGGwZD;vdkBVVkZvzsG= NUKwXWVWOTZ{NkexNlQ>
PC 12 NFTaPGJHfW6ldHnvckBCe3OjeR?= Mo\HNVDDqM7:TR?= MWKyOEBp MYnBeJRmdnWjdHXzJINifGWlaH;sZY1qdmViYnnvd5lvfGinc3nz MUexOlIyQTR{NB?=
Cynomolgus monkey embryonic stem cells MWLDfZRwfG:6aXOgRZN{[Xl? MVyyNEDDvU1? Mn;4NlQhcA>? MWrQdo9ud3SnczDjfWVUKGOnbHygd5Vzfmm4YXy= MXGxPFk1ODh3NR?=
TSGH 8301 NXmwT4xsVWmpcnH0bY9vKEG|c3H5 M1LNV|IxKML3TR?= MX6xJIg> M3;wUWlv[3KnYYPld{Bk\WyuIH3p[5JifGmxbh?= NGKyOVIyQTh7NkS3OS=>
Swiss3T3 NXrzS4l5S2:ub375MYZwem2rbnegRZN{[Xl? MorZNVAhyrWP MoD0NVMh\A>? NGjtPXFKdmO{ZXHz[ZMheHKxc4TheIUh[2WubDDjc4xwdnlvZn;ycYlv\yCjY4Tpeol1gQ>? NFPWdlczOTR4NEmwNi=>
HT22 MnjjR5l1d3SxeHnjJGF{e2G7 MoW3NVAhyrWP MW[xN{Bp NGK3WlVRem:2ZXP0d{Bi\2GrboP0JIdtfXSjbXH0[U1qdmS3Y3XkJI5mfXKxbnHsJIRm[XSq Mk\TNlI5OTB6M{W=
Salivary gland stem cells NFe2UY9HfW6ldHnvckBCe3OjeR?= Mo\JNVAhyrWP MnXXO{Bl M2nNVnJm\HWlZYOgV2dUSyC|ZX7ld4NmdmOn MnvkNlU5ODR3NkC=

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western Blot
p-LIMK1(Thr508); 

PubMed: 24704720     


Immunoblot analysis of HCT116 and HCT116 SMAD4-/- cells. Cells were transfected transiently with either BMPR2 or pcDNA vector. Subsequently, cells were treated with 2.5 μmol/L of ROCK inhibitor (Y-27632) or PBS. Blots were incubated with antibodies against pLIMK1/2 and p-cofilin. Actin was used as a loading control.

p-LIMK2(Thr505); 

PubMed: 24704720     


Immunoblot analysis of HCT116 and HCT116 SMAD4-/- cells. Cells were transfected transiently with either BMPR2 or pcDNA vector. Subsequently, cells were treated with 2.5 μmol/L of ROCK inhibitor (Y-27632) or PBS. Blots were incubated with antibodies against pLIMK1/2 and p-cofilin. Actin was used as a loading control.

p-Cofilin(Ser3); 

PubMed: 24704720     


Immunoblot analysis of HCT116 and HCT116 SMAD4-/- cells. Cells were transfected transiently with either BMPR2 or pcDNA vector. Subsequently, cells were treated with 2.5 μmol/L of ROCK inhibitor (Y-27632) or PBS. Blots were incubated with antibodies against pLIMK1/2 and p-cofilin. Actin was used as a loading control.

CDK2; 

PubMed: 26555939     


Western blot detected proteins changes pTR cells cultured with Y-27632. CDX2 was remarkably increased in Y-27632 treated pIVFTR-7 and pPATR-5 cells. pTR cells cultured without Y-27632 in the same condition are used as control. β-ACTIN serves as an endogenous control. 

KRT7; 

PubMed: 26555939     


Western blot detected proteins changes pTR cells cultured with Y-27632. CDX2 was remarkably increased in Y-27632 treated pIVFTR-7 and pPATR-5 cells. pTR cells cultured without Y-27632 in the same condition are used as control. β-ACTIN serves as an endogenous control. 

ROCK2; 

PubMed: 26555939     


Western blot detected proteins changes pTR cells cultured with Y-27632. CDX2 was remarkably increased in Y-27632 treated pIVFTR-7 and pPATR-5 cells. pTR cells cultured without Y-27632 in the same condition are used as control. β-ACTIN serves as an endogenous control. 

E-cadherin; 

PubMed: 27399334     


The expression and/or phosphorylation of E-cadherin and ROCKs downstream targets in SW-480-FOXM1D cells treated with Y-27632.

p-MLC2(Ser19); 

PubMed: 27399334     


The expression and/or phosphorylation of E-cadherin and ROCKs downstream targets in SW-480-FOXM1D cells treated with Y-27632.

24704720 26555939 27399334
Transwell migration assay
cell migration inhibition ; 

PubMed: 27694793     


Data represent mean ± SEM, n=3. Compared with control, * P<0.05, ** P<0.01. Compared with control, Y-27632, or EGCG, *** P<0.01.

27694793
Immunofluorescence
Neurotoxicity Assay; 

PubMed: 21362567     


Representations of neurodegeneration in H9-, HUF5- and G2019S-iPSC derived neurons when treated with neurotoxins, H2O2 and MG-132, and ROCK inhibitor Y-27632. Scale bar = 50 µm.

E-cadherin; 

PubMed: 24523903     


The localized patterns of E-cadherin and b-catenin were examined using specific antibodies through confocal laser microscopy. MCF-7 cells were cultured for 24 hrs in the absence or presence of Y-27632 (20 µM) to inhibit ROCK activity. 

beta-catenin; 

PubMed: 24523903     


The localized patterns of E-cadherin and b-catenin were examined using specific antibodies through confocal laser microscopy. MCF-7 cells were cultured for 24 hrs in the absence or presence of Y-27632 (20 µM) to inhibit ROCK activity. 

Phalloidin; 

PubMed: 27399334     


Triple IF staining for F-actin (phalloidin, green) E-cadherin (b) (red) and nuclei (DAPI, blue) showed decreased F-actin, increased E-cadherin and altered cell shape and size in SW-480-FOXM1D cells treated with the ROCKs inhibitor Y-27632 or fasudil (both 10 μM, for 24 h). Scale bar, 25 μm. 

21362567 24523903 27399334
Growth inhibition assay
cell proliferation ; 

PubMed: 24523903     


MCF-7 cells and MDA-MB-231 cells were cultured on tissue culture plates for four days with or without Y-27632 (20 µM). Relative cell proliferation rates were determined by the cell number assayed using CCK-8 kit. The data are expressed as the mean ± SD. n = 3 culture dishes. The p-value was less than 0.05 for comparisons between control (Control) and treatment groups (Y-27632) in MCF-7 cells. *, p<0.05.

24523903
ELISA
caspase-9; 

PubMed: 30320378     


Caspase-9 activity was measured via ELISA. Mst1-mediated casapse-9 activation was inhibited by Y-27632 in A549 cells. *P<0.05. Ad, adenovirus; Mst1, mammalian STE20-like kinase 1.

30320378
In vivo Oral administration of Y-27632 2HCl at 30 mg/kg significantly decreases the blood pressure in a dose-dependent manner in spontaneous hypertensive rats, renal hypertensive rats, as well as deoxycorticosterone acetate (DOCA)-salt hypertensive rats. [1] When Y-27632 2HCl is continuously administered at a rate of 0.55 μL per hour by implanted pumps for 11 days tumor cell invasion (MM1 cells expressing Val14-RhoA in rats) is significantly delayed. [2] By inhibiting ROCK, Y-27632 2HCl treatment attenuates hypoxia-induced angiogenesis and vascular remodeling in the pulmonary circulation. [5] Pretreatment with Y-27632 has a protective effect against tumor formation in albino mice with Ehrlich ascites carcinoma. [7]

Protocol

Animal Research:[1] [7]
- Collapse
  • Animal Models: Male Wistar rats with spontaneous or induced hypertension; Swiss albino mice with Ehrlich ascites carcinoma
  • Dosages: 30 mg/kg/day (Rat); 0-10 mg/kg (mice)
  • Administration: Orally (Rat); i.p. (Mice)
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 64 mg/mL warmed (199.83 mM)
Water 14 mg/mL (43.71 mM)
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
saline
For best results, use promptly after mixing.
10mg/mL

* 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.

Chemical Information

Molecular Weight 320.26
Formula

C14H21N3O.2HCl

CAS No. 129830-38-2
Storage powder
in solvent
Synonyms N/A
Smiles Cl.Cl.CC(N)C1CCC(CC1)C(=O)NC2=CC=NC=C2

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

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Frequently Asked Questions

  • Question 1:

    Is there any data about the Amax (maximum attraction luminosity) and extinction coefficient of this compound?

  • Answer:

    The wavelength we used to test HPLC is 260nm while the extinction coefficient is unknown.

  • Question 2:

    Could this product be used in cell culture? Do you have any reference for this application?

  • Answer:

    Yes. The Y-27632 can be used in cell culture certainly. Here is the reference website: http://molpharm.aspetjournals.org/content/57/5/976.full.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID