VAS2870-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEVAS2870Cat.No.:HY-12804CASNo.:722456-31-7分⼦式:C₁₈H₁₂N₆OS分⼦量:360.39作⽤靶点:NADPHOxidase作⽤通路:MetabolicEnzyme/Protease储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:83.3mg/mL(231.14mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.7748mL13.8739mL27.7477mL5mM0.5550mL2.7748mL5.5495mL10mM0.2775mL1.3874mL2.7748mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。体内实验请根据您的实验动物和给药⽅式选择适当的溶解⽅案。以下溶解⽅案都请先按照InVitro⽅式配制澄的储备液，再依次添加助溶剂：(为保证实验结果的可靠性，澄的储备液可以根据储存条件，适当保存；体内实验的⼯作液，建议您现⽤现配，当天使⽤；以下溶剂前显⽰的百分⽐指该溶剂在您配制终溶液中的体积占⽐；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的⽅式助溶)1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(6.94mM);Clearsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE2.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.94mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性VAS2870NADPH氧化酶(NADPHoxidase(NOX))抑制剂。IC50&TargetTarget:NADPHoxidase[1]体外研究VAS2870iseffectivetosuppressPDGF-BB-dependentactivationofNADPHoxidaseandsubsequentproductionofintracellularROS.Furthermore,VAS2870suppressesPDGF-BB-dependentchemotaxis,butnotDNAsynthesis.PreincubationwithVAS2870(10and20μM)completelyabolishesPDGF-mediatedNADPHoxidaseactivationandROSproduction.PreincubationwithVAS2870(0.1-20μM)doesnotaffectPDGF-inducedcellcycleprogression.However,itabolishesPDGF-dependentchemotaxisofVSMCinaconcentration-dependentmanner(100%inhibitionat10μM)[1].VAS2870inhibitsdose-dependentlyautocrineincreaseofcellnumberinFaOrathepatomacells,andalmostcompletelyblockedROSproductionandthymidineincorporationwhenusedat25mM.VAS2870blocksserum-dependentcellgrowthofFaOrathepatomacells.VAS2870inhibitsproliferationofdifferenthumanhepatocellularcarcinoma(HCC)celllines.VAS2870pretreatmentenhancesTGF-b-mediatedapoptosisofFaOrathepatomacells[2].PROTOCOLKinaseAssay[1]NADPHoxidaseactivityismeasuredbylucigenin-enhancedchemiluminescenceina50mMphosphatebuffer(bufferA),pH7.0,containing1mMEGTA,proteaseinhibitors,150mMsucrose,5μMlucigenin,and250μMNADPHassubstrate.Quiescentcellsarestarvedbyserumdeprivationfor24handtreatedasindicated,ishedtwicewithice-coldphosphatebufferedsaline(PBS),pH7.4,andharvested.Afterlowspincentrifugation,thepelletisre-suspendedinice-coldbufferA,lackinglucigeninandsubstrate.Then,thecellsarelysedandtotalproteinconcentrationisdeterminedusingaBradfordassayandadjustedto1mg/mL.100μLaliquotsoftheproteinsamplearemeasuredover6mininquadruplicatesusingNADPH(100μM)assubstrateinascintillationcounter.Dataarecollectedat2minintervalsinordertomeasurerelativechangesinNADPHoxidaseactivity[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]Totestautocrinegrowth,cellsareserumdeprivedat40%confluenceand,whenindicated,theNADPHoxidaseinhibitorsApocynin(300mM)orVAS2870areadded30minbeforeserumdeprivationandmaintainedalongtheexperiment.ForTGF-bexperiments,cellsat70%confluenceareserumdeprivedfor16handtreatedwith2ng/mLTGF-βinthepresenceorabsenceoftheEGFRinhibitorAG1478(20mM)orVAS2870(25mM),whichareadded30minpriortoTGF-β[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE•FreeRadicBiolMed.2022Feb24;S0891-5849(22)00080-6.•FreeRadicBiolMed.2022Mar;181:166-179.•OxidMedCellLongev.27Feb2022.•MolCellEndocrinol.2020Dec1;518:110990.•BiochemBiophysResCommun.2020Apr9;524(3):575-581.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].tenFreyhausH,etal.NovelNoxinhibitorVAS2870attenuatesPDGF-dependentsmoothmusclecellchemotaxis,butnotproliferation.CardiovascRes.2006Jul15;71(2):331-41.[2].SanchoP,etal.TheNADPHoxidaseinhibitorVAS2870impairscellgrowthandenhancesTGF-β-inducedapoptosisoflivertumorcells.BiochemPharmacol.2011Apr1;81