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Medchemexpress/BIX-01294/HY-10587/10mM*1mL in DMSO
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Medchemexpress/BIX-01294/HY-10587/10mM*1mL in DMSO
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BIX-01294isaninhibitorofG9aHistoneMethyltransferasewithIC50of1.9μM.

CustomerValidation

  • CellDeathDis.2017Apr6;8(4):e2726.
  • HarvardMedicalSchoolLINCSLIBRARY
Description

BIX-01294isaninhibitorofG9aHistoneMethyltransferasewithIC50of1.9μM.

IC50&Target

IC50:1.9μM(G9aHistoneMethyltransferase)

InVitro

BIX-01294(4.1μM)treatmentof G9anullEScellsresultsinafurtherreductioninH3K9me2levels.BIX-01294reducesH3K9me2atseveralG9atargetgenes[1].BIX-01294(5µM)reduceslevelsofglobalH3K9methylationinpronuclear-and2-cell-stageembryos.EmbryostransientlyexposedtoBIX-01294haveareducedABIlitytoestablishpregnancy[2].BIX-01294(1µg/mL)causesreductionintheBrdUincorporationoffetalPASMCs.BIX-01294treatmentdecreasesthePASMCsmigrationinducedbyPDGF[3].

References
  • [1].KubicekS,etal.ReversalofH3K9me2byasmall-moleculeinhibitorfortheG9ahistonemethyltransferase.MolCell.2007Feb9;25(3):473-81.

    [2].ParkKE,etal.IVMBIX-01294,aninhibitorofthehistonemethyltransferaseEHMT2,disruptshistoneH3lysine9(H3K9)dimethylationinthecleavage-stageporcineembryo.ReprodFertilDev.2012;24(6):813-21.

    [3].YangQ,etal.BIX-01294treatmentblockscellproliferation,migrationandcontractilityinovinefoetalpulmonaryarterialsmoothmusclecells.CellProlif.2012Aug;45(4):335-44.

PreparingStockSolutions
ConcentrationVolumeMass1mg5mg10mg
1mM2.0382mL10.1908mL20.3815mL
5mM0.4076mL2.0382mL4.0763mL
10mM0.2038mL1.0191mL2.0382mL
Pleaserefertothesolubilityinformationtoselecttheappropriatesolvent.
KinaseAssay
[1]

Testcompoundsaredilutedto12μg/mLin50mMTris-HClpH8.5containing4%DMSOand10μLisdispensedintothewells.Blankandcontrolwellsreceiveonlycompoundbuffer.GST-G9aat10μg/mLandSAMat40μMaredilutedin50mMTrisHClpH8.5/10mMDTTandaddedinavolumeof20μL.BlankwellsreceiveTris/DTTbufferonly.Thereactionsareinitiatedbytheadditionof800nMH3(1-20)-cySBIotinsubstratein50mMTrispH8.5inavolumeof10μL,andincubatedatroomtemperaturefor60minutes.Theplatesarewashed3timeswith100μLofishBuffer(50mMTrispH7.4,150mMNaCl,0.05%Tween20,0.2%BSA).Next,50μLofluoroimmunoassay(FI)Buffer(50mMTrisHClpH7.8,150mMNaCl,0.05%Tween40,25μMDTPA,0.2%BSA,0.05%BGG)containing5ngα-2X-di-methH3-K9and5nggoatanti-rabbitEuchelateisaddedtoallwellsoftheplate,andtheplateisincubatedforanadditionalhouratroomtemperature.Theplatesarewashed3timeswith100μLofishBuffer,and50μLofEnhancementSolutionisaddedtoeachwell.Timeresolvedfluorescenceismeasuredafter45minutesonaViewluxMicroplateImagerimagingfor15secondswitha354μswindow,400μsdelay,excitationat360nm,andemissionat618nm.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

CellAssay
[3]

Briefly,fetalPASMCsareplatedin96-wellplatesandstarvedfor24hat0.1%serumcontainingmedium.PDGF-BBisaddedfor24hattheindicatedconcentrationsinthepresenceorabsenceofBIX-01294.BrdUlabelsolutionisaddedtoeachwell18hpriortoanalysis.Denaturingsolutionisaddedtoeachwellfor30minatroomtemperatureafterremovingthecontentsofwells.Then,anti-BrdUantibodyisaddedtoeachwellandincubatedfor1handperoxidasegoatanti-mouseIgGHRPconjugateisaddedinthewellfor30minatroomtemperature.Theabsorbanceisreadat450-540nmonaGlomaxMultipleDetectionSystem.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

References
  • [1].KubicekS,etal.ReversalofH3K9me2byasmall-moleculeinhibitorfortheG9ahistonemethyltransferase.MolCell.2007Feb9;25(3):473-81.

    [2].ParkKE,etal.IVMBIX-01294,aninhibitorofthehistonemethyltransferaseEHMT2,disruptshistoneH3lysine9(H3K9)dimethylationinthecleavage-stageporcineembryo.ReprodFertilDev.2012;24(6):813-21.

    [3].YangQ,etal.BIX-01294treatmentblockscellproliferation,migrationandcontractilityinovinefoetalpulmonaryarterialsmoothmusclecells.CellProlif.2012Aug;45(4):335-44.

MolecularWeight

490.64

Formula

C₂₈H₃₈N₆O₂

CASNo.

935693-62-2

Storage
Powder-20°C3years
 4°C2years
Insolvent-80°C6months
 -20°C1month
Shipping

RoomtemperatureincontinentalUS;mayvaryelsewhere

Solvent&Solubility

10mMinDMSO

*"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">

Purity:99.22%