Description | MG-132isapotent,non-specific20Sproteasomeinhibitor,withIC50of24.2nMfortheβ5chymotrypsin-likeactivesite. |
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IC50&Target | IC50:24.2nM(chymotrypsin-likeactivity)[1] |
InVitro | Dose-dependentinhibitionofcellgrowthisobservedinHeLacellswithanIC50ofapproximately5μMMG132for24h.MG132inhibitsthegrowthofHeLacellsviainducingthecellcyclearrestaswellastriggeringapoptosis[2].MG-132inhibitsC6gliomacellproliferationinatime-anddose-dependentmanner(theIC50valueat24his18.5μM).MG-132(18.5μM)suppressestheproteasomeactivitybyabout70%at3h.MG-132inducesapoptosisviadown-regulationofantiapoptoticproteinsBcl-2andXIAP,up-regulationofpro-apoptoticproteinBaxandcaspase-3,andproductionofcleavedC-terminal85kDaPARP.MG-132alsocausesamorethan5-foldincreaseofreactiveoxygenspecies[3].TheIC50ofMG-132againstHeLa,CaSki,andC33AcervicalcancercellsviABIlityafter48hofincubationis2.1,3.2,and5.2μM,respectively[4]. |
InVivo | TheinvivoantitumoractivityofMG-132againstcervicalcancerisexaminedusings.c.xenograftmodels.MG-132isinjectedat1mg/kgusingthefollowingschedule:days1,4,8,12,1518,23,and26formicebearingHeLatumors.ThegrowthinhibitionratesofMG132comparedtocontrolis49%[4].MG-132(i.p.,0.1mg/kg/day)attenuatespressure-overload-inducedcardiachypertrophyandimprovescardiacfunctioninaBDominalaorticbanding(AAB)ratsthroughregulationofERK1/2andJNK1signalingpathways[5]. |
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PreparingStockSolutions |
Pleaserefertothesolubilityinformationtoselecttheappropriatesolvent. | ||||||||||||||||
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KinaseAssay [3] | Aftergrowingonsix-wellplates(3×105cells/well)for24h,C6gliomacellsaretreatedwitheitherPBS(control)or18.5μMMG-132for3,6,12,or24hat37°C.CellsarethoroughlyscrapedfromtheculturedisheswithacellscraperandwashedwithcoldPBS.Aftercentrifugationfor10minat800×g,thecellpelletsaresUSPendedinice-coldbuffer(50mMTris-HCl,pH7.5,20μMATP,5mMMgCl2,1mMdithiothreitol,and20%glycerol)andhomogenizedwithaPyrexglassmicrohomogenizer(20strokes).Thehomogenateiscentrifugedat15000×gfor10minat4°Ctoobtainsupernatant.Proteinconcentrationisdeterminedusingproteinassaykits.Atotalof10μL(1μg/μL)ofeachfreshlymadesupernatantisincubatedina96-wellplateat37°Cfor30minwith10μLof300μMofSuccinyl-LLVY-AMCand85μLofassaybuffer(20mMTris-HCl,pH7.5,and20%glycerol).ReleaseoffluorescentAMCismeasuredwithaspectrofluorometerat440nmwithanexcitationwavelengthof380nm[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly. | ||||||||||||||||
CellAssay [3] | MG-132isdissolvedinPBStoastorageconcentrationof50μM[3]. C6gliomacellsareseededonto96-wellmicroplates(3×104cells/well)andculturedfor24h.ThecellsaretreatedwithPBSorMG-132finalconcentrationsof10,20,30,and40μM,respectively.CellviabilityisassessedusinganMTTassayat3,6,12,and24hafterMG-132treatment.Theabsorbancevalueat570nmisreadusinganautomaticmulti-wellspectrophotometer.C6gliomacells(3×105cells/well)areallowedtogrowoncoverslipsin6-wellcultureplatesfor24h.ThecellsarethentreatedwitheitherPBS(control)or18.5μMMG-132at37°Cfor24h.Cellsgrowingonglasscoverslipsarefixedinmethanolfor5minatroomtemperature.ThefixedcellsarewashedtwicewithPBSandthenincubatedwithHoechst33342for5minatroomtemperatureandobservedunderafluorescencemicroscope.Fragmentedorcondensednucleiarescoredasapoptotic[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly. | ||||||||||||||||
AnimalAdmiNISTration [4][5] | MG-132isdissolvedinvehicle(saline)(Mice)[4]. Mice[4] | ||||||||||||||||
References |
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MolecularWeight | 475.62 | |
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Formula | C₂₆H₄₁N₃O₅ | |
CASNo. | 133407-82-6 | |
Storage | 4°C,protectfromlight | |
Shipping | RoomtemperatureincontinentalUS;mayvaryelsewhere | |
Solvent&Solubility | 10mMinDMSO *"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">1> Purity:>98.0%
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