Rhodamine-123-solution-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemERhodamine123(solution)Cat.No.:HY-DY1054CASNo.:62669-70-9分⼦式:C₂₁H₁₇ClN₂O₃分⼦量:380.83作⽤靶点:FluorescentDye作⽤通路:Others储存⽅式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY⽣物活性罗丹明类染料⼀种膜渗透性的阳离⼦荧光探针，可对线粒体膜电位进⾏特异性识别，从⽽附在线粒体中，并产⽣明亮荧光，在⼀定浓度下，罗丹明类染料对细胞具有低毒性，因此遍⽤于检测动物细胞、植物细胞以及微⽣物中的线粒体。溶剂及浓度：DMSO：5mM体外研究1.PreparationofRhodamine123workingsolution1.1PreparationofthestocksolutionDissolve1mgRhodamine123in525μLDMSOtoobtain5mMofstocksolution.1.2PreparationofRhodamine123workingsolutionDilutethestocksolutioninserum-freecellculturemedium.Thecorrespondingstocksolutioncanbedilutedaccordingtotheactualsituation.NotethatifthesolventisDMSO,thecytotoxicityofDMSOmustbeconsidered,andasolventcontrolshouldbeprepared;ifthesolventispurewater,theworkingsolutionneedstobefilteredandsterilizedbeforeaddingcells.Note:PleaseadjusttheconcentrationofRhodamine123workingsolutionaccordingtotheactualsituation.2.Cellstaining2.1Suspensioncells(6-wellplate)a.Centrifugeat1000gat4°Cfor3-5minutesandthendiscardthesupernatant.WashtwicewithPBS,5minuteseachtime.Thecelldensityis1×106/mL.b.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,andthenincubateatroomtemperaturefor30-60minutes.d.Washtwicewithmedium,5minuteseachtime.Observationbyfluorescencemicroscopyorflowcytometry.Note:Ifdetectionbyflowcytometry,cellsneedtoberesuspendedbeforestaining.REFERENCES[1].M.Huang,etal.MitochondrialInnerMembraneElectrophysiologyAssessedbyRhodamine-123TransportandFluorescence.AnnBiomedEng.2007Jul;35(7):1276–1285.[2].Emaus,R.K.,Grunwald,R.,&Lemasters,J.J.(1986).Rhodamine123asaprobeoftransmembranepotentialinisolatedrat-livermitochondria:spectralandmetabolicproperties.BiochimicaetBiophysicaActa(BBA)-Bioenergetics,850(3),436–448.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedicalapplication