FM4-64-solution-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEFM4-64(solution)Cat.No.:HY-DY1031CASNo.:162112-35-8分⼦式:C₃₀H₄₅Br₂N₃分⼦量:607.52作⽤靶点:FluorescentDye作⽤通路:Others储存⽅式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY⽣物活性FM4-64(solution)⼀种亲脂性很强的⽔溶性苯⼄烯染料，能特异性结合细胞膜和内膜细胞器产⽣荧光。FM4-64⼴泛⽤于内吞和外显膜结构标记。溶剂及浓度：DMSO：5mM体外研究1.PreparationofFM4-64workingsolution1.1StocksolutionItisrecommendedtostorethestocksolutionat-20°Cor-80°Cawayfromlightandavoidrepetitivefreeze-thawcycles.1.2PreparationofFM4-64workingsolutionDilutethestocksolutioninHBSS.Thecorrespondingstocksolutioncanbedilutedaccordingtotheactualsituation.NotethatifthesolventisDMSO,thecytotoxicityofDMSOmustbeconsidered,andasolventcontrolshouldbeprepared;ifthesolventispurewater,theworkingsolutionneedstobefilteredandsterilizedbeforeaddingcells.Note:PleaseadjusttheconcentrationofFMworkingsolutionaccordingtotheactualsituation.2.Cellstaining2.1Suspensioncells(6-wellplate)a.Centrifugeat1000gat4°Cfor3-5minutesandthendiscardthesupernatant.WashtwicewithPBS,5minuteseachtime.Thecelldensityis1×106/mLb.Add1mLofworkingsolution,andthenincubateatroomtemperaturefor5-30minutes.c.Centrifugeat400gat4°Cfor3-4minutesandthendiscardthesupernatant.d.WashtwicewithPBS,5minuteseachtime.e.Resuspendcellswithserum-freecellculturemediumorPBS.Observationbyfluorescencemicroscopyor1/2MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEflowcytometry.2.2Adherentcellsa.Cultureadherentcellsonsterilecoverslips.b.Removethecoverslipfromthemediumandaspirateexcessmedium.c.Add100μLofworkingsolution,gentlyshakeittocompletelycoverthecells,andthenincubateatroomtemperaturefor5-30minutes.d.Washtwicewithmedium,5minuteseachtime.Observationbyfluorescencemicroscopyorflowcytometry.StorageStoreinseparatecontainerstoavoidrepeatedfreeze-thawcycles.-20°C,1year.Protectfromlight.REFERENCES[1].ZalT,etal.SpectralshiftoffluorescentdyeFM4-64revealsdistinctmicroenvironmentofnuclearenvelopeinlivingcells.Traffic.2006;7(12):1607-1613.[2].StenovecM,etal.DistinctlabellingoffusioneventsinratlactotrophsbyFM1-43andFM4-64isassociatedwithconformationaldifferences.ActaPhysiol(Oxf).2007;191(1):35-42.[3].BolteS,etal,Satiat-JeunemaitreB.FM-dyesasexperimentalprobesfordissectingvesicletraffickinginlivingplantcells.JMicrosc.2004;214(Pt2):159-173.[4].JEGale,etal.FM1-43dyebehavesasapermeantblockerofthehair-cellmechanotransducerchannel.JNeurosci.2001Sep15;21(18):7013-25.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedicala