Dihydrorhodamine-123-solution-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEDihydrorhodamine123(solution)Cat.No.:HY-DY1037CASNo.:109244-58-8分⼦式:C₂₁H₁₈N₂O₃分⼦量:346.39作⽤靶点:FluorescentDye作⽤通路:Others储存⽅式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY⽣物活性Dihydrorhodamine123(DHR123)(solution)⼀种⾮荧光活性氧(ROS)指⽰剂。Dihydrorhodamine123在细胞内存在活性氧物质的情况下被氧化为荧光试剂Rhodamine123(HY-D0816)，并定位于线粒体中(Ex/Em=515/536nm)。溶剂及浓度：DMSO：10mM体外研究1.Solutionpreparation1.1StocksolutionStorage:Storeat-20°Cor-80°Cindarkafteraliquoting.Avoidrepeatedfreezingandthawing.1.2PreparationofworkingsolutionDilutewithcellculturemedium(optimizedaccordingtotheexperiment).Thecorrespondingstocksolutioncanbedilutedaccordingtotheactualsituation.NotethatifthesolventisDMSO,thecytotoxicityofDMSOmustbeconsidered,andasolventcontrolshouldbeprepared;ifthesolventispurewater,theworkingsolutionneedstobefilteredandsterilizedbeforeaddingcells.Note:Theworkingsolutionshouldbepreparedandusedimmediately.Keepitawayfromlight.2.Cellstaining2.1Treatcellswithinducersorinhibitorsofreactiveoxygenspecies.Note:Iftreatingcellswithoxidizingagents,rinsecellsthreetimeswithbufferorculturemediumtoremovethecompoundsbeforestainingtoavoiddirectoxidationofthedyebythecompound.2.2DiluteDihydrorhodamine123dyestocksolutioninserum-freeculturemediumatafinalconcentrationof5μM.2.3Removemediumfromthecellsandreplacewithmediumcontainingdye.1/2MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE2.4Incubatefor30minutesat37°C.2.5Detectbyfluorescencemicroscopyorflowcytometry.Washingisnotrequired,butcanbeperformedtoreducebackgroundformicroscopy.Inthepresenceof10μMDihydrorhodamine123(DHR123)thestimulationoftheneutrophilNADPHoxidasebytheadditionof50nMphorbol12-myristate13-acetat(PMA)resultesinanincreaseintherateofrhodaminegeneration.Thefluorescentintensityofthecells,inthepresenceof10μMDihydrorhodamine123,increaseswithtimefollowingtheadditionof50nMPMA.Inthepresenceof10μMDihydrorhodamine123,inducedHL60cellsshowasustainedincreaseinfluorescencefollowingtheadditionof50nMPMA[1].REFERENCES[1].LydiaM.Hendersonetal.Dihydrorhodamine123:afluorescentprobeforsuperoxidegeneration?Eur.J.Biochem.217,973-980.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedicalapplications.For