Y-27632 2HCl

For research use only.

Catalog No.S1049

494 publications

Y-27632 2HCl Chemical Structure

CAS No. 129830-38-2

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 494 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 NFXWVYxHfW6ldHnvckBCe3OjeR?= MmLCNVAh|ryP M4TuN|IhcA>? M4XZUWROW09? M164RmlvcGmkaYTzJJRp\SCjc4PlcYJtgSCxZjDtbYNzd3S3YoXs[ZMh[W6mIHnueIVzdWWmaXH0[UBncWyjbXXueJMhfG9iZn;ycUBmgHSnbnTl[EBxem:lZYPz[ZM> NV7zdHZXQTZ2N{[1OC=>
N1E-115 M17vNGZ2dmO2aX;uJGF{e2G7 M{\4UFExKM7:TR?= MnyzNkBp MX3EUXNQ MYrJcohq[mm2czD0bIUh[XO|ZX3icJkhd2ZibXnjdo91fWK3bHXzJIFv\CCrboTldo1m\GmjdHWg[olt[W2nboTzJJRwKG[xcn2g[Zh1\W6mZXSgdJJw[2W|c3Xz NHX6UZk6PjR5NkW0
HeLa MmX1SpVv[3Srb36gRZN{[Xl? M2TOSVExKM7:TR?= Mn6yN|AhdWmw M3ruN2lvcGmkaYTzJJRp\SCob4LtZZRqd25ib3[gd5Rz\XO|IH\pZoVzeyCjbnSgeIhmKGG|c3XtZox6KG:oII\pcoN2dGmwLXPvcpRicW6rbneg[o9k[WxiYXTo[ZNqd26| MVG5OlY5ODd{
CCL39 NYS2UHdVTnWwY4Tpc44hSXO|YYm= NV;SXI96OzBizszN M1XQU|MxKG2rbh?= MVTDc41xdGW2ZXz5JIFjd2yrc3jld{Bi[3SrdnH0bY9vKG:oIF7hMWgh\XilaHHu[4VzKE6KRUGgZpkhcW62ZXfybY5{ MoK2PVY6OzN6Mh?=
Mesothelial cells from rat mesentery MX3JcpZie2m4ZTDBd5NigQ>? Mo\KN|Ah|ryP M4n0NFIxKGh? MkLsRoxw[2u|IHnueoF{cX[nIHHjeIl3cXS7 MWK5PVMxQDd{
NIH3T3 MY\Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NVnRVXFmOTBizszN MX6xPEBl NIO1RmRFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp MXSxNFAzOTN6Nh?=
Dbl-d NETq[5VIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NYXRe2dHOTBizszN MnzBNVgh\A>? M2iyTnN1em:wZ3z5JIlvcGmkaYTzJINmdGxiZ4Lve5Rp MYKxNFAzOTN6Nh?=
Dbl-e M13oOGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NWXJ[W1uOTBizszN M1fseVE5KGR? Mn;OUY9l\XKjdHXsfUBqdmirYnn0d{Bk\WyuIHfyc5d1cA>? M4Dr[lExODJzM{i2
mNET1-d MWrHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MnmyNVAh|ryP NHXMdWoyQCCm M1:4WHN1em:wZ3z5JIlvcGmkaYTzJINmdGxiZ4Lve5Rp M1zIdlExODJzM{i2
mNET1-e NV\icYFoT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MoPFNVAh|ryP NWmzO2IyOThiZB?= MWfTeJJwdmeueTDpcohq[mm2czDj[YxtKGe{b4f0bC=> NXm3bZdPOTByMkGzPFY>
Ras-2 MnHaS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NWrpSWlQOTBizszN M4PBO|E5KGR? MnXuV5Rzd26pbImgbY5pcWKrdIOgZ4VtdCCpcn;3eIg> MUSxNFAzOTN6Nh?=
Ras-4 NF7l[YxIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NUWxemVsOTBizszN NWLHcXdoOThiZB?= MmCzV5Rzd26pbImgbY5pcWKrdIOgZ4VtdCCpcn;3eIg> NFXUZ4EyODB{MUO4Oi=>
Src-1 MmjES5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? Mmi1NVAh|ryP M{D2[FE5KGR? NFTtN4NFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp MnnlNVAxOjF|OE[=
Src-4 MXHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NWDVTVNbOTBizszN NFS2ZWcyQCCm M3nyV2Rw\XNibn;0JIlvcGmkaYSgZ4VtdCCpcn;3eIg> MWqxNFAzOTN6Nh?=
NIH3T3 M2\hcmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MnfsNVAh|ryP NE\XbIQyQCCm M2j5T2Rw\XNibn;0JIlvcGmkaYSgZ4VtdCCpcn;3eIg> NFvWZpkyODB{MUO4Oi=>
Src-1 Mn7ES5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NYK4WlJWOTBizszN MkLnNVgh\A>? NH;X[nBFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp M1jCOlExODJzM{i2
Src-2 M2fYNWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MnXtNVAh|ryP MnzpNVgh\A>? NUjEenFjTG:nczDuc5QhcW6qaXLpeEBk\WyuIHfyc5d1cA>? M4LDO|ExODJzM{i2
SW620 NFfPVZNIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NFXoZXkyOCEQvF2= MnvLNVgh\A>? MnXtSI9meyCwb4SgbY5pcWKrdDDj[YxtKGe{b4f0bC=> MXyxNFAzOTN6Nh?=
HCT15 Mn\LS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NIPRNFAyOCEQvF2= NEHKXlUyQCCm NGC3VZVFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp MYmxNFAzOTN6Nh?=
HCT116 MVnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MWexNEDPxE1? NH;6co0yQCCm Mom5V5Rzd26pbImgbY5pcWKrdIOgZ4VtdCCpcn;3eIg> M4myW|ExODJzM{i2
LS174T NF\POWZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M{LPZVExKM7:TR?= M{LvWVE5KGR? MV7Nc4RmemG2ZXz5JIlvcGmkaYTzJINmdGxiZ4Lve5Rp NELrcXEyODB{MUO4Oi=>
Neonatal rat ventricular myocytes MWrGeY5kfGmxbjDBd5NigQ>? M4ji[VExKM7:TR?= NXHtWnUxPDhiaB?= M3zGbWlvcGmkaYTzJGVVNTFvaX7keYNm\CCrbnPy[YF{\XNiaX6gdJJwfGWrbjDzfY51cGW|aYOsJINmdGxic3n6[UBidmRibYnv[oljemmubHHyJI9z\2GwaYrheIlwdg>? NHvV[FQyODN6Nk[xNy=>
Stellate Cell NIL2Ro9HfW6ldHnvckBCe3OjeR?= NFPM[HYzPSEQvF2= MWixOUBucW5? NWL6SVhLUW6qaXLpeJMh\m:{bXH0bY9vKG:oIF[tZYN1cW5ic4Ty[ZN{KG[rYnXyd{BidmRicHjvd5Bpd3K7bHH0bY9vKG:oIH35c5NqdiCuaXfoeEBkcGGrbh?= NGGzRlkyODZyMES5Oi=>
Rat Vascular Smooth Muscle Cells MUfGeY5kfGmxbjDBd5NigQ>? NXniUXpOOTBizszN MlnBNkBp Mo\rTY5pcWKrdIOgZY5ocW:2ZX7zbY4hUUlvaX7keYNm\CCqeYDldpRzd3CqeR?= MV[xNFY1OjNzNx?=
PC3 NITSbIpHfW6ldHnvckBCe3OjeR?= MX:yOUDPxE1? M3u2dlEhcA>? NYfWRWJHUW6mdXPld{Bud3KyaH;sc4dq[2GuIHPoZY5o\XN? NYC0eplpOTB5MkC0O|E>
PC3 NXe2Vo9kVWmpcnH0bY9vKEG|c3H5 MnHNNlUh|ryP MlHYNUBp NVThVoJ6UW6qaXLpeJMhfGinIFLNSmIuS01iYX7kJJRp\SCHR1[td5RqdXWuYYTl[EBucWe{YYTpc44> NW[1VG1jOTB5MkC0O|E>
PC3 NIfJOYNIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NUj1dYFYOjVizszN MVuxO{Bp Ml7pSI9meyCwb4SgbY5pcWKrdDDj[YxtKGe{b4f0bC=> NVvGS5JCOTB5MkC0O|E>
LNCaP NYDvZYtiT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MlfoNlUh|ryP Mlm3NVchcA>? MnPLSI9meyCwb4SgbY5pcWKrdDDj[YxtKGe{b4f0bC=> NYjy[5ZKOTB5MkC0O|E>
Rat hepatic stellate cells NVfrO3Q{TnWwY4Tpc44hSXO|YYm= MkDNN|Ah|ryP M4XRO|Q5KGh? MUfEbY1qdmm|aHXzJJRp\SCyaH;zdIhwenmuYYTpc44hd2ZiRYLrNkwh[W6mIHTlZ5Jm[XOnczDu[ZchTE6DIIP5cpRp\XOrcx?= NEjYcYgyODh2NU[2Ny=>
Pancreatic acinar cells MVXGeY5kfGmxbjDBd5NigQ>? NFviZ40yOMLizszN MnLhO|AhdWmw NX\TcVlwWG:2ZX70bYF1\XNiQ1PLMZN1cW23bHH0[YQheGGwY4LlZZRq[yCnbor5cYUhe2WlcnX0bY9v NF\rVHAyOjd2NUC4NC=>
C2C12 MU\GeY5kfGmxbjDBd5NigQ>? MnzINVDDqM7:TR?= M4PwNlYhcA>? M4Pw[3Bz\X[nboTzJJRp\SC|ZYLpcoUheGixc4Doc5J6dGG2aX;uJI9nKEmUUz2xJIlv\HWlZXSgZpkhcW6|dXzpckBidmRxb4KgWG5HNc7z NVTTTJNzOTZ{NkexNlQ>
PC 12 NX\4V4xoTnWwY4Tpc44hSXO|YYm= Ml;mNVDDqM7:TR?= M{DkbVI1KGh? NVzSd3ZESXS2ZX71ZZRmeyClYYTlZ4hwdGGvaX7lJIJqd3O7boTo[ZNqew>? NI[3UnoyPjJzOUSyOC=>
Cynomolgus monkey embryonic stem cells MYjDfZRwfG:6aXOgRZN{[Xl? NG\pO5EzOCEEtV2= MkTvNlQhcA>? MWnQdo9ud3SnczDjfWVUKGOnbHygd5Vzfmm4YXy= MXKxPFk1ODh3NR?=
TSGH 8301 NHjGdW1OcWe{YYTpc44hSXO|YYm= MkGyNlAhyrWP NXXqT|RGOSCq MXTJcoNz\WG|ZYOgZ4VtdCCvaXfyZZRqd25? NIjMTpMyQTh7NkS3OS=>
Swiss3T3 M3rmU2NwdG:weT3mc5JucW6pIFHzd4F6 MYCxNEDDvU1? MmDFNVMh\A>? M1jqV2lv[3KnYYPld{Bxem:|dHH0[UBk\WyuIHPvcI9vgS2ob4LtbY5oKGGldHn2bZR6 MWSyNVQ3PDlyMh?=
HT22 NYe4c|I4S3m2b4TvfIlkKEG|c3H5 NULYW3pxOTBiwsXN NFy5[ZkyOyCq M2XweXBzd3SnY4TzJIFo[Wmwc4Sg[4x2fGGvYYTlMYlv\HWlZXSgcoV2em:wYXyg[IVifGh? M1q4XVIzQDFyOEO1
Salivary gland stem cells NHjoV|FHfW6ldHnvckBCe3OjeR?= NEfkbGoyOCEEtV2= MlS2O{Bl NHTkbY1T\WS3Y3XzJHNIW0Nic3Xu[ZNk\W6lZR?= NFHKWoczPThyNEW2NC=>

... 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 CC(C1CCC(CC1)C(=O)NC2=CC=NC=C2)N.Cl.Cl

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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