H2DCFDA-solution-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEH2DCFDA(solution)Cat.No.:HY-DY1002作⽤靶点:ReactiveOxygenSpecies(ROS);FluorescentDye作⽤通路:Immunology/Inflammation;MetabolicEnzyme/Protease;NF-κB;Others储存⽅式:-20°C,protectfromlight.Avoidrepeatedfreezingandthawing*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light.Avoidrepeatedfreezingandthawing)BIOLOGICALACTIVITY⽣物活性H2DCFDA(DCFH-DA)(solution)⼀种可渗透细胞的，⽤于检测细胞内活性氧(ROS)的探针(Ex/Em=488/525nm)[1]。溶剂及浓度：DMSO：10mM体外研究Guide(Thefollowingisourrecommendedprotocol.Thisprotocolisonlyaguideandshouldbemodifiedaccordingtoyourspecificneeds).1Thecorrespondingstocksolutioncanbedilutedaccordingtotheactualsituation.NotethatifthesolventisDMSO,thecytotoxicityofDMSOmustbeconsidered,andasolventcontrolshouldbeprepared;ifthesolventispurewater,theworkingsolutionneedstobefilteredandsterilizedbeforeaddingcells.2.Cellstaining(suspendedcells)2.1CollectcellsbycentrifugationandwashtwicewithPBSfor5minuteseachtime.Thecelldensityis1×106/mL.2.2Add1mLofdyeworkingsolutionandincubateatroomtemperaturefor5-30minutes.2.3Centrifugeat400gfor3-4minutesanddiscardthesupernatant.2.4AddPBStowashthecellstwice,5minuteseachtime.2.5Resuspendthecellsin1mLofserum-freemediumorPBSandobserveusingafluorescencemicroscopeorflowcytometer.3.Cellstaining(adherentcells)3.1Culturetheadherentcellsonasterilecoverslip.3.2Removethecoverslipfromtheculturemediumandremovetheexcessculturemedium.3.3Add100μLofdyeworkingsolution,gentlyshaketocompletelycoverthecells,andincubatefor5-30minutes.3.4Aspiratethedyeworkingsolution,wash2-3timeswithculturemedium,5minuteseachtime,and1/2MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEobserveusingafluorescencemicroscopeorflowcytometer.Note:Ifflowcytometryisrequired,thecellsneedtobedigestedwithtrypsinandresuspendedbeforestaining.REFERENCES[1].LyublinskayaOG,etal.Redoxenvironmentinstemanddifferentiatedcells:Aquantitativeapproach.RedoxBiol.2017Aug;12:758-769.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedicalapplications.Forresearchuse