H2DCFDA-DCFH-DA-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEH2DCFDACat.No.:HY-D0940CASNo.:4091-99-0Synonyms:DCFH-DA;2',7'-Dichlorodihydrofluoresceindiacetate分⼦式:C₂₄H₁₆Cl₂O₇分⼦量:487.29作⽤靶点:ReactiveOxygenSpecies作⽤通路:Immunology/Inflammation;MetabolicEnzyme/Protease;NF-κB储存⽅式:-20°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light)溶解性数据体外实验DMSO:125mg/mL(256.52mM;Needultrasonic)Ethanol:20mg/mL(41.04mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.0522mL10.2608mL20.5217mL5mM0.4104mL2.0522mL4.1043mL10mM0.2052mL1.0261mL2.0522mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month(protectfromlight)。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。体内实验请根据您的实验动物和给药⽅式选择适当的溶解⽅案。以下溶解⽅案都请先按照InVitro⽅式配制澄的储备液，再依次添加助溶剂：(为保证实验结果的可靠性，澄的储备液可以根据储存条件，适当保存；体内实验的⼯作液，建议您现⽤现配，当天使⽤；以下溶剂前显⽰的百分⽐指该溶剂在您配制终溶液中的体积占⽐；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的⽅式助溶)1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESolubility:2.08mg/mL(4.27mM);Suspendedsolution;Needultrasonic2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.27mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.27mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性H2DCFDA(DCFH-DA)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]。体外研究Guidelines(Followingisourrecommendedprotocol.Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds).1.H2DCFDAisdissolvedinDMSOtoobtaina10mMstocksolutionsandfurtherdilutedbeforeuse.2.Cellsareincubatedwith5µMH2DCFDAsolutioninPBSinthedarkfor30minat37°C,thenharvestedwith0.05%trypsin-EDTAsolution,suspendedinafreshmedium,andimmediatelyanalyzedwithflowcytometer.3.AlongwiththeH2DCFDAprobe,ifindicated,useROS-insensitivemodificationofthefluoresceindyeDCFDAasapositivecontrol.ThestainingprocedureisthesameasfortheH2DCFDA[1].PROTOCOLKinaseAssayROSMeasurements[1]ForthedetectionofintracellularROSlevel,ROS-sensitiveprobeH2DCFDAisused.Adherentcells(ESCs,difESCs,eMSCs,HeLa,U118)areincubatedwith5µMstainingsolutioninPBSinthedarkfor30minat37°C,thenharvestedwith0.05%trypsin-EDTAsolution,suspendedinafreshmedium,andimmediatelyanalyzedwithflowcytometer.Lymphocytes,bothcontrolandPHA-activated,areresuspendedinPBS,incubatedwith5µMofH2DCFDAinthedarkfor30minat37°C,andimmediatelyanalyzed.AlongwiththeH2DCFDAprobe,ifindicated,ROS-insensitivemodificationofthefluorescentdyeDCFDAisused.ThestainingprocedureisthesameasfortheH2DCFDA[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•Cell.2021Jun24;184(13):3528-3541.e12.•SignalTransductTargetTher.2022Aug15;7(1):288.•NanoEnergy.2022:108002.•ACSNano.2022Oct11.•ACSNano.2021Oct26;15(10):16286-16297.Seemorecustomervalidationsonwww.MedChemEREFERENCES2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE[1].LyublinskayaOG,etal.Redoxenvironmentinstemanddifferentiatedcells:Aquantitativeapproach.RedoxBiol.2017Aug;12:758-769.McePdfHeightCaution:Producthasnotbeen