VAS2870-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEVAS2870Cat.No.:HY-12804CASNo.:722456-31-7分?式:C??H??N?OS分?量:360.39作?靶點:NADPHOxidase作?通路:MetabolicEnzyme/Protease儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:83.3mg/mL(231.14mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.7748mL13.8739mL27.7477mL5mM0.5550mL2.7748mL5.5495mL10mM0.2775mL1.3874mL2.7748mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內使?，-20°C儲存時，請在1個?內使?。體內實驗請根據(jù)您的實驗動物和給藥?式選擇適當?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實驗結果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當保存；體內實驗的?作液，建議您現(xiàn)?現(xiàn)配，當天使?；以下溶劑前顯?的百分?指該溶劑在您配制終溶液中的體積占?；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶)1.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(6.94mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.94mM);ClearsolutionBIOLOGICALACTIVITY?物活性VAS2870NADPH氧化酶(NADPHoxidase(NOX))抑制劑。IC50&TargetTarget:NADPHoxidase[1]體外研究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.戶使?本產(chǎn)品發(fā)表的科研?獻2/3MasterofBioactiveMolecules—您?邊的抑制劑?師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