For research use only.

Catalog No.S1102

604 publications

U0126-EtOH Chemical Structure

CAS No. 1173097-76-1

U0126-EtOH is a highly selective inhibitor of MEK1/2 with IC50 of 0.07 μM/0.06 μM in cell-free assays, 100-fold higher affinity for ΔN3-S218E/S222D MEK than PD98059. U0126 inhibits autophagy and mitophagy with antiviral activity.

Selleck's U0126-EtOH has been cited by 604 publications

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Biological Activity

Description U0126-EtOH is a highly selective inhibitor of MEK1/2 with IC50 of 0.07 μM/0.06 μM in cell-free assays, 100-fold higher affinity for ΔN3-S218E/S222D MEK than PD98059. U0126 inhibits autophagy and mitophagy with antiviral activity.
Features A chemically synthesized and highly selective inhibitor of both MEK1 and MEK2.
MEK2 [1]
(Cell-free assay)
MEK1 [1]
(Cell-free assay)
0.06 μM 0.07 μM
In vitro

U0126-EtOH functionally antagonizes AP- 1 transcriptional activity and blocks the production of a variety of cytokines and metalloproteinases involved in the inflammatory response. [1] U0126-EtOH inhibits T cell proliferation in response to antigenic stimulation or cross-linked anti-CD3 plus anti-CD28 Abs without effect on IL-2-induced proliferation by down-regulating IL-2 mRNA levels. [2] A recent study shows that U0126-EtOH antagonizes resveratrol-induced apoptosis in castration-resistant human prostate cancer C4-2 cells, inhibits mitochondrial function and shifts cells to aerobic glycolysis independently of MEK. [3]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
rat PC12 cells NUDifoJYTnWwY4Tpc44h[XO|YYm= MWKxNEDPxE1? MWCxJIg> M1Xpc2FkfGm4YYTpc44hd2ZiToLmNk9CWkViaX6gdoF1KFCFMUKgZ4VtdHNiYYPz[ZN{\WRiYYOgTG8uOSCycn;0[YlvKGmwZIXjeIlwdiCjdDCxNEB2VSCjZoTldkA2KGi{czDwdoV1emWjdHXkJJdqfGhiSl7LJIlvcGmkaYTvdkBUWDZyMEGyOUBnd3JiMTDodkBj\W[xcnWgZ49ueG:3bnSgZYRlcXSrb36gZpkhX2W|dHXyckBjdG:2IHHuZYx6e2m| NWjtWIZGRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkGzOFU3QDVpPkKxN|Q2Pjh3PD;hQi=>
mouse RAS-3T3 cells NXrPcmhYTnWwY4Tpc44h[XO|YYm= NE\iTGEyOC12MDFOwG0> NGGzXJJKdmirYnn0bY9vKG:oIF3FT{1u\WSrYYTl[EBGWktzL{KgdIhwe3Cqb4L5cIF1cW:wIHnuJI1wfXOnIGLBV{0{XDNiY3XscJMh[XRiMUCgeI8hPDBidV2gZpkhTUyLU1Gu NV[2UpdORGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS1NFc5OjZpPkK0OVA4QDJ4PD;hQi=>
HCT116 cells NF7sPFBHfW6ldHnvckBie3OjeR?= NUPSV4loSWKrbHn0fUBw\iClb33wc5Vv\CC2bzDpcohq[mm2IHHuZ4hwemGpZTDpcoRmeGWwZHXueEBkd2yxbomg[o9zdWG2aX;uJEh{d2[2IHHnZZIh\3Kxd4ToJIF{e2G7KTDpckBJS1RzMU[gZ4VtdHNuIFnDOVA:OTlwNDFOwG0v MX68ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPTJ{NUewOkc,OTV{MkW3NFY9N2F-
COS-7 cells NH;YR2NHfW6ldHnvckBie3OjeR?= NH7tXnFKdmirYnn0c5J6KGOxbnPlcpRz[XSrb36gZYdicW6|dDDBVE0yKHS{YX7zZ5JqeHSrb36gbY4hS0:VLUegZ4VtdHQxvJygTWM2OD1zIN88UU4> MV[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPTByNkO4Okc,OTVyME[zPFY9N2F-
HeLa cells NUX4V5JJTnWwY4Tpc44h[XO|YYm= MXzJcohq[mm2aX;uJI9nKEWJRj3zeIlufWyjdHXkJGVtczFvbIXjbYZmemG|ZTDy[ZBwenSncjDhd5NigSCrbjDI[WxiKGOnbHzzMEBKSzVyPUCuNlkh|ryPLh?= NWjVdWVCRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUWyNlU4ODZpPkG1NlI2PzB4PD;hQi=>

... Click to View More Cell Line Experimental Data

Methods Test Index PMID
Western blot
p-MEK / MEK / p-ERK / ERK; 

PubMed: 27487136     

Representative blots of MEK1/2 and ERK1/2, either total or phosphorylated, in A673 and TC71 cell lines treated with vehicle (DMSO) or 20 μM U0126. 

E-cadherin / Vimentin / Twist-2; 

PubMed: 28416770     

Western blot for E-cadherin, Vimentin, Twist-2, phoshpo-ERK1/2 and total ERK1/2 in total cell lysates of OVCAR-3-NID1-MC cells after U0126 treatment. GAPDH served as an internal control of protein loading.

27487136 28416770
pERK / pPPARγ; 

PubMed: 27145370     

Decreased phosphorylated ERK1/2 (red) and phosphorylated PPARγ (green) with U0126 treatment in high glucose condition (30 mM). 

E-cadherin / Vimentin; 

PubMed: 28416770     

Immunofluorescent microscopic images of E-cadherin (red) and Vimentin (red) in OVCAR-3-NID1-MC cells after U0126 treatment. Nuclear was stained with 4,6-diamidino-2-phenylindole (blue) (scale bar = 20 μm).


PubMed: 26828592     

LB39-MEL CD70+ cells were treated with 5 μM of U0126 for 72 h then fixed and processed for immunofluorescence directed against CD70. Pictures of U0126 treated cells and control (DMSO) conditions are presented (scale bar 50 μm) 

27145370 28416770 26828592
In vivo U0126-EtOH, as the inhibitor of intracellular Raf/MEK/ERK signaling pathway, demonstrates antiviral activity by suppressing propagation of the 2009 pandemic IV H1N1 variant and highly pathogenic avian influenza viruses (HPAIV) in vivo in the mouse lung by inhibiting. [4] U0126-EtOH shows the potential neuroprotective effect and improving spatial learning in Morris water maze (MWM) by activating peroxisome proliferator-activated receptor gamma coactivator-1a, nuclear respiratory factor 1, and mitochondrial transcription factor A in Aβ-injected rats. [5]


Kinase Assay:


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In Vitro Kinase Assays :

The amount of immunoprecipitated wild type MEK used in these assays is adjusted to give a similar amount of activity units as obtained with 10 nM recombinant MEK. Reaction velocities are measured using a 96-well nitrocellulose filter apparatus as described below. Unless otherwise noted, reactions are carried out at an enzyme concentration of 10 nM, in 20 mM Hepes, 10 mM MgCl2, 5 mM β-mercaptoethanol, 0.1 mg/mL BSA, pH 7.4, at room temperature. Reactions are initiated by the addition of [γ-33P]ATP into the premixed MEK/ERK/inhibitor reaction mixture, and an aliquot of 100 μL is taken every 6 minutes and transferred to the 96-well nitrocellulose membrane plate which has 50 mM EDTA to stop the reaction. The membrane plate is drawn and washed 4 times with buffer under vacuum. Wells are then filled with 30 μL of Microscint-20 scintillation fluid, and the radioactivity of 33P-phosphorylated ERK is counted with a Top Count scintillation counter. Velocities are obtained from the slopes of radioactivity versus time plots. Concentrations of ERK and ATP are 400 nM and 40 μM, respectively, unless otherwise indicated. For all of the in vitro enzyme assays, the percent inhibition is calculated 100 (1 −Vi/Vo) where Vi and Vo are the initial reaction velocities in the presence and absence of inhibitor, respectively. The data are then plotted as percent inhibition as a function of inhibitor concentration and fit, by nonlinear least squares regression, to the standard equation for a Langmuir isotherm to determine the IC50. As reported, enzyme concentrations are based upon molecular weights and mass of protein used in the final assay volume and not on active site titration. Thus, the actual enzyme active site concentration may differ from that reported.
Cell Research:


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  • Cell lines: A.E7 or Th17 cells
  • Concentrations: 0 to 10 μM
  • Incubation Time: 48 hours
  • Method:

    A.E7 or Th17 cells are incubated with mitomycin C-treated B10.BR or BALB/c splenocytes plus varying concentrations of pigeon cytochrome c or PR8 Ag, or with 5 U/mL human rIL-2. In addition, some assays contains U0126 or an inactive analogue, U0124, to determine direct effects of MEK inhibition on T cell proliferation. Two days after culture initiation, each well is pulsed with 1 µCi of [3H]TdR and harvested the following day. The incorporation of [3H]TdR into DNA is quantitated on a Packard Matrix 96 direct beta counter without the use of liquid scintillation mixtures.

    (Only for Reference)
Animal Research:


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  • Animal Models: Female C57Bl/6 mice infected by Mouse-adapted highly pathogenic avian influenza A/FPV/Bratislava/79 (H7N7; FPV) virus and swine origin human influenza A virus (SOIV) A/Regensburg/D6/2009 (H1N1v; RB1).
  • Dosages: ≤10 mM
  • Administration: Administered via aerosol.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 85 mg/mL (199.26 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
5%DMSO+40%PEG300+5%Tween 80+50%H2O
For best results, use promptly after mixing.

* 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 426.56


CAS No. 1173097-76-1
Storage powder
in solvent
Synonyms N/A
Smiles CCO.C1=CC=C(C(=C1)N)SC(=C(C#N)C(=C(N)SC2=CC=CC=C2N)C#N)N

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
Dosage mg/kg Average weight of animals g Dosing volume per animal ul Number of animals
Step 2: Enter the in vivo formulation ()
% DMSO % % Tween 80 % ddH2O

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This equation is commonly abbreviated as: C1V1 = C2V2 ( Input Output )

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* When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and SDS / COA (available online).

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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

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Frequently Asked Questions

  • Question 1:

    I want to know whether the compound is light-sensitive?

  • Answer:

    S1102 U0126-EtOH is not stable. It should be stored as powder at -20°C, and prepared the solution just before use.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID