Y-27632 2HCl

Catalog No.S1049

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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Cited by 322 Publications

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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 MXfGeY5kfGmxbjDBd5NigQ>? NHjXT4EyOCEQvF2= M{jWXlIhcA>? NHPKb2NFVVOR NH\BSm1KdmirYnn0d{B1cGViYYPz[Y1jdHlib3[gcYlkem:2dXL1cIV{KGGwZDDpcpRmem2nZHnheIUh\mmuYX3lcpR{KHSxIH\vdo0h\Xi2ZX7k[YQheHKxY3Xzd4V{ MXy5OlQ4PjV2
N1E-115 M3;q[mZ2dmO2aX;uJGF{e2G7 MXqxNEDPxE1? NHX0NoozKGh? NV7vS|JDTE2VTx?= NFz6TItKdmirYnn0d{B1cGViYYPz[Y1jdHlib3[gcYlkem:2dXL1cIV{KGGwZDDpcpRmem2nZHnheIUh\mmuYX3lcpR{KHSxIH\vdo0h\Xi2ZX7k[YQheHKxY3Xzd4V{ NHHKeIc6PjR5NkW0
HeLa NW\yemJCTnWwY4Tpc44hSXO|YYm= M2DQNFExKM7:TR?= MWOzNEBucW5? NUSwWG11UW6qaXLpeJMhfGinIH\vdo1ifGmxbjDv[kB{fHKnc4Og[olj\XK|IHHu[EB1cGViYYPz[Y1jdHlib3[geolv[3WuaX6tZ49vfGGrbnnu[{Bnd2OjbDDh[Ihme2mxboO= NWP3PHFCQTZ4OEC3Ni=>
CCL39 NIfaWWFHfW6ldHnvckBCe3OjeR?= MnrTN|Ah|ryP MUWzNEBucW5? MV3Dc41xdGW2ZXz5JIFjd2yrc3jld{Bi[3SrdnH0bY9vKG:oIF7hMWgh\XilaHHu[4VzKE6KRUGgZpkhcW62ZXfybY5{ MofaPVY6OzN6Mh?=
Mesothelial cells from rat mesentery NYrWXG1[UW64YYPpeoUhSXO|YYm= NGLxWoY{OCEQvF2= NYnWV5VrOjBiaB?= M1LDVWJtd2OtczDpcpZie2m4ZTDhZ5Rqfmm2eR?= MmfyPVk{ODh5Mh?=
NIH3T3 M3jscGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NIjvUJIyOCEQvF2= M4\3eVE5KGR? NWjS[VV1TG:nczDuc5QhcW6qaXLpeEBk\WyuIHfyc5d1cA>? NFnMbW8yODB{MUO4Oi=>
Dbl-d NYjDcm1DT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NEXEc4syOCEQvF2= NX73RlZtOThiZB?= NWXqWnplW3S{b37ncJkhcW6qaXLpeJMh[2WubDDndo94fGh? MVSxNFAzOTN6Nh?=
Dbl-e NETpe4ZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MXixNEDPxE1? MnzCNVgh\A>? NID4cmdOd2SncnH0[Yx6KGmwaHnibZR{KGOnbHyg[5Jwf3Sq M33NZ|ExODJzM{i2
mNET1-d MXfHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NXH1[FdOOTBizszN MWexPEBl Ml;PV5Rzd26pbImgbY5pcWKrdIOgZ4VtdCCpcn;3eIg> MorHNVAxOjF|OE[=
mNET1-e M{TrOGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M3\WO|ExKM7:TR?= MmHoNVgh\A>? M1rNXXN1em:wZ3z5JIlvcGmkaYTzJINmdGxiZ4Lve5Rp NVO3UHJjOTByMkGzPFY>
Ras-2 NHHYNWhIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NY\Cdo9LOTBizszN M2r1R|E5KGR? NE\ydJFUfHKxbnfsfUBqdmirYnn0d{Bk\WyuIHfyc5d1cA>? NXfpUIJ[OTByMkGzPFY>
Ras-4 NHzLPHZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NX3qe3k{OTBizszN MVyxPEBl NVvBNoM6W3S{b37ncJkhcW6qaXLpeJMh[2WubDDndo94fGh? MmrsNVAxOjF|OE[=
Src-1 M3PnSWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MkGwNVAh|ryP NYLVRXFROThiZB?= NELFc5RFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp NHG1bJQyODB{MUO4Oi=>
Src-4 MWLHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MXexNEDPxE1? M3XBOVE5KGR? MUHEc4V{KG6xdDDpcohq[mm2IHPlcIwh\3Kxd4To NYfYNHp2OTByMkGzPFY>
NIH3T3 MVzHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M{nUb|ExKM7:TR?= MnjsNVgh\A>? NFjBUWtFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp NXmxTWVpOTByMkGzPFY>
Src-1 MXTHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NH3pWJoyOCEQvF2= NFXwbZcyQCCm M1v3dmRw\XNibn;0JIlvcGmkaYSgZ4VtdCCpcn;3eIg> MYmxNFAzOTN6Nh?=
Src-2 MXnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M4HtNVExKM7:TR?= M3fGe|E5KGR? Mo[4SI9meyCwb4SgbY5pcWKrdDDj[YxtKGe{b4f0bC=> M3;nflExODJzM{i2
SW620 M1;LNGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M3S1RVExKM7:TR?= NXfrRVRROThiZB?= MYDEc4V{KG6xdDDpcohq[mm2IHPlcIwh\3Kxd4To NUPWXnJpOTByMkGzPFY>
HCT15 MWnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MkG3NVAh|ryP M{fnRlE5KGR? NXPUXZp[TG:nczDuc5QhcW6qaXLpeEBk\WyuIHfyc5d1cA>? MoTpNVAxOjF|OE[=
HCT116 MnLzS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M4jyeVExKM7:TR?= NV\lfpI{OThiZB?= Mn\VV5Rzd26pbImgbY5pcWKrdIOgZ4VtdCCpcn;3eIg> NG\ObVMyODB{MUO4Oi=>
LS174T MmPwS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MY[xNEDPxE1? Mn7INVgh\A>? M3PtUm1w\GW{YYTlcJkhcW6qaXLpeJMh[2WubDDndo94fGh? Ml23NVAxOjF|OE[=
Neonatal rat ventricular myocytes M4TSSmZ2dmO2aX;uJGF{e2G7 NHXwTWQyOCEQvF2= NXLncYdyPDhiaB?= NWLJUoVTUW6qaXLpeJMhTVRvMT3pcoR2[2WmIHnuZ5Jm[XOnczDpckBxem:2ZXnuJJN6dnSqZYPpd{wh[2WubDDzbZpmKGGwZDDtfY9ncWK{aXzsZZIhd3KpYX7pfoF1cW:w NH;CSJoyODN6Nk[xNy=>
Stellate Cell M3rhZWZ2dmO2aX;uJGF{e2G7 NUnDdXpQOjVizszN NWHCNGdHOTVibXnu MYrJcohq[mm2czDmc5Ju[XSrb36gc4YhTi2jY4TpckB{fHKnc4Og[olj\XK|IHHu[EBxcG:|cHjvdplt[XSrb36gc4YhdXmxc3nuJIxq\2i2IHPoZYlv NWm3UZVJOTB4MEC0PVY>
Rat Vascular Smooth Muscle Cells NY[3dVBCTnWwY4Tpc44hSXO|YYm= MWWxNEDPxE1? Mnj0NkBp M4ntWmlvcGmkaYTzJIFv\2mxdHXud4lvKEmLLXnu[JVk\WRiaInw[ZJ1em:yaIm= MoXCNVA3PDJ|MUe=
PC3 NUTiZXBuTnWwY4Tpc44hSXO|YYm= NVTiXFROOjVizszN MUmxJIg> Mme1TY5lfWOnczDtc5JxcG:ub3fpZ4FtKGOqYX7n[ZM> MXyxNFczODR5MR?=
PC3 M3L3[21q\3KjdHnvckBCe3OjeR?= NVOzR4d[OjVizszN NYjJRppVOSCq MYnJcohq[mm2czD0bIUhSk2IQj3DUUBidmRidHjlJGVITi2|dHnteYxifGWmIH3p[5JifGmxbh?= MnPHNVA4OjB2N{G=
PC3 NXKweXluT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MVeyOUDPxE1? NUPubYtXOTdiaB?= NGDIWmtFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp MmH1NVA4OjB2N{G=
LNCaP M4DOUGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NEjkOWMzPSEQvF2= MWWxO{Bp NG\EfZdFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp Mn\oNVA4OjB2N{G=
Rat hepatic stellate cells MVPGeY5kfGmxbjDBd5NigQ>? NUTjfmJoOzBizszN MYK0PEBp NYDZRYVYTGmvaX7pd4hmeyC2aHWgdIhwe3Cqb4L5cIF1cW:wIH;mJGVzczJuIHHu[EBl\WO{ZXHz[ZMhdmW5IFTORUB{gW62aHXzbZM> NUL1[lhIOTB6NEW2OlM>
Pancreatic acinar cells MlvGSpVv[3Srb36gRZN{[Xl? M{fPXFExyqEQvF2= MVi3NEBucW5? NIjsdmZRd3SnboTpZZRmeyCFQ1utd5RqdXWuYYTl[EBx[W6lcnXheIlkKGWweont[UB{\WO{ZYTpc44> MWqxNlc1PTB6MB?=
C2C12 MWfGeY5kfGmxbjDBd5NigQ>? MlXhNVDDqM7:TR?= NWHpO4syPiCq NFf1OlRRemW4ZX70d{B1cGVic3XybY5mKHCqb4PwbI9zgWyjdHnvckBw\iCLUmOtNUBqdmS3Y3XkJIJ6KGmwc4XsbY4h[W6mL3;yJHRPTi4QsR?= M3LONlE3OjZ5MUK0
PC 12 M4T1NGZ2dmO2aX;uJGF{e2G7 M2rkNFExyqEQvF2= MVGyOEBp M3;hV2F1fGWwdXH0[ZMh[2G2ZXPoc4xidWmwZTDibY9{gW62aHXzbZM> NUDWdG5UOTZ{MUm0NlQ>
Cynomolgus monkey embryonic stem cells NInOcJREgXSxdH;4bYMhSXO|YYm= MlT1NlAhyrWP NEnqe4QzPCCq MoKyVJJwdW:2ZYOgZ5lGWyClZXzsJJN2en[rdnHs NY\XT3VSOTh7NEC4OVU>
TSGH 8301 NVKybo5OVWmpcnH0bY9vKEG|c3H5 MmjmNlAhyrWP M4H2SlEhcA>? Mmm1TY5kemWjc3XzJINmdGxibXnndoF1cW:w NHr0dHIyQTh7NkS3OS=>
Swiss3T3 M{PwN2NwdG:weT3mc5JucW6pIFHzd4F6 M2XBSFExKML3TR?= MljoNVMh\A>? NEHzOpJKdmO{ZXHz[ZMheHKxc4TheIUh[2WubDDjc4xwdnlvZn;ycYlv\yCjY4Tpeol1gQ>? NETvNYwzOTR4NEmwNi=>
HT22 MnTUR5l1d3SxeHnjJGF{e2G7 NGPHVYMyOCEEtV2= NGLsR3oyOyCq MXrQdo91\WO2czDh[4FqdnO2IHfseZRidWG2ZT3pcoR2[2WmIH7leZJwdmGuIHTlZZRp M4T0S|IzQDFyOEO1
Salivary gland stem cells MmLRSpVv[3Srb36gRZN{[Xl? MU[xNEDDvU1? MUW3JIQ> M17KfHJm\HWlZYOgV2dUSyC|ZX7ld4NmdmOn M{nZRVI2QDB2NU[w

... 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]
+ Expand
  • Animal Models: Male Wistar rats with spontaneous or induced hypertension; Swiss albino mice with Ehrlich ascites carcinoma
  • Formulation: Dissolved in DMSO, and diluted in saline (Rat); 0.9% NaCl (Mice)
  • 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

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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