JC-1-solution-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEJC-1(solution)Cat.No.:HY-DY1003CASNo.:3520-43-2分⼦式:C₂₅H₂₇Cl₄IN₄分⼦量:652.22作⽤靶点:FluorescentDye作⽤通路:Others储存⽅式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY⽣物活性JC-1(CBIC2)(solution)⼀种⼴泛⽤于检测线粒体膜电位的理想荧光探针。JC-1染料以电势依赖性的⽅式积聚在线粒体内，可以⽤来检测细胞、组织或纯化的线粒体膜电位。正常线粒体内，JC-1聚集在线粒体质中形成聚合物，聚合物发出强烈的红⾊荧光(Ex=585nm,Em=590nm)；在线粒体膜电位较低时，JC-1不能聚集在线粒体的质中，产⽣绿⾊荧光(Ex=510nm,Em=527nm)。溶剂及浓度：DMSO：1.5mM体外研究PreparationofJC-1workingsolution:1‰DMSO+89.9%ddH2O+10%10xPBS(HY-K3006).Preparea1.5-30μMJC-1workingsolution.Take1mLofthe1.5μMworkingsolutionasanexample:Remove1μLfromthe1.5mMJC-1(solution)andaddittothetube.Mixwellwith899μLofddH2O,thenadd100μLof10xPBStotheabovemixtureandmixthoroughly.Note:1)TheJC-1dyeneedstobepreparedimmediatelyandusedassoonaspossible.Itmayprecipitateifleftforaperiodoftime;trytoprepareanduseitunderalight-proofcondition.2)Ateachstepofthedilutionprocess,ensurethoroughmixingtoachieveclarity.Ifnecessary,ultrasonicassistancefordissolutionat37℃canbeused.3)Pleasedonotdirectlydiluteitwith1xPBS(HY-K3005)orserum-freemediumtoobtaintheworkingsolution;otherwise,itwillleadtosevereprecipitation.4)IftheeffectofJC-1inenteringthecellsisnotsatisfactory,anappropriateamountof20%PluronicF127solutioncanbeaddedtotheworkingsolution,withafinalconcentrationof0.02-0.05%.PluronicF127canpreventJC-1fromaggregatinginthebuffersolutionandhelpitenterthecells.5)ThestoragesolutionofJC-1isrecommendedtobealiquotedandstoredat-20°Cor-80°Cinthedark.1/2MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEJC-1Staining:a.Takethe6-wellplateasanexampleforcellplanking,andthedensityis5×105/mL.Incubateovernightin5%CO2incubatorat37℃.Note::Itissuggestedthatthecelldensityduringapoptosisinductionshouldnotexceed1×106/ml,whichcanalsobeculturedtotheappropriatedensityaccordingtoyourowncelltype.b.Transfer0.5mLofthecellsuspensionintoasterilecentrifugetube.c.Take0.5mLsuspensionintosterilecentrifugetube;400gcentrifugationfor3-5min;Discardthesupernatant.d.Thecellswereresuspendedwith1mLJC-1workingsolutionandincubatedin5%CO2incubatorat37℃for15-30min.e.Centrifugationatroomtemperaturefor5minat400g;Suckofthesupernatant.f.Thecellswereresuspendedwith2mLcellculturemediumorbuffer,andthencentrifugedatroomtemperaturefor5minat400g;Discardthesupernatantandrepeattwice.g.Resuspendthecellswith1mLoffreshculturemediumorbuffer,andimmediatelyconductsubsequentflowcytometryorfluorescencemicroscopeobservation.h.Dataanalysis(flowcytometry):mitochondriaofhealthycellscontainingredJC-1aggregatesweredetectedbyFL2channel;ApoptoticorunhealthycellscontaininggreenJC-1monomerweredetectedbyFL1(FITC)channel.Note:Ifusedforenzymelabelinginstrument,use300μLbufferresuspendedcells;Then,transferthestainedcellstoanopaque96-wellplateatarateof100μLperwell,andthenconductfluorescentenzymelabelplateanalysis.REFERENCES[1].APerelman,etal.JC-1:alternativeexcitationwavelengthsfacilitatemitochondrialmembranepotentialcytometry.CellDeathDis.2012Nov22;3:e430.[2].VeraC.Keil,etal.Ratiometrichigh-resolutionimagingofJC-1fluorescencerevealsthesubcellularheterogeneityofastrocyticmitochondria.PflügersArchiv-EuropeanJournalofPhysiology.2011,462(5):693-708.[3].Jung-HoLEE,etal.Real-timeanalysisofamyloidfibrilformationofα-synucleinusingafibrillation-state-specificfluorescentprobeofJC-1.Biochem.J.2009,418:311-323.[4].SalvioliS,etal.JC-1,butnotDiOC6(3)orrhodamine123,isareliablefluorescentprobetoassessdeltapsichangesinintactcells:implicationsforstudiesonmitochondrialfunctionalityduringapoptosis.FEBSLett.1997Jul7;411(1):77-82.Mce