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Biotium/NucView 530 Caspase-3 Substrate, 1 mM in DMSO/10406-T/trial-size-10-ul
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NucView™530Caspase-3Substrate,1mMinDMSO,providesaconvenienttoolfordetectingapoptosisinintactcellsbasedoncaspase-3/7activityusingeitherconfocalmicroscopyorflowcytometry.NucView™530Caspase-3Substrate,1mMinDMSO,providesaconvenienttoolfordetectingapoptosisinintactcellsbasedoncaspase-3/7activityusingeitherconfocalmicroscopyorflowcytometry.Incontrasttootherfluorogeniccaspasesubstratesorfluorescentcaspaseinhibitorbased(FLICA)assays,NucView™caspase-3substratescanbeusedtodetectcaspase-3/7activitywithinindividualintactcellswithoutinhibitingapoptosisprogression.NucView™substratesconsistofafluorogenicDNAdyecoupledtothecaspase-3/7DEVDrecognitionsequence.Thesubstrate,whichisinitiallynon-fluorescent,penetratestheplasmamembraneandentersthecytoplasm.Inapoptoticcells,caspase-3/7cleavesthesubstrate,releasingthehigh-affinityDNAdye,whichmigratestothecellnucleusandstainsDNAwithfluorescence.Thus,NucView™Caspase-3Substratesarebifunctional,allowingdetectionofcaspase-3/7activityandvisualizationofmorphologicalchangesinthenucleusduringapoptosis.Thestainingisalsoformaldehyde-fixable.NucView™530Caspase-3Substratestainsapoptoticcellnucleiwithorangefluorescence(Ex/Em528/563nm,Figure1),fordetectionintheCy®3channelbyfluorescencemicroscopy(Figure2)orthePEchannelbyflowcytometry(Figure3).NucView™530canbeusedformulti-colorimagingwithblue,green,orfar-redfluorescentprobes.Notethatwhenexcitedbythe488nmlaserline,NucView™530alsofluorescesintheFITCchannel,andthereforecannotbeanalyzedtogetherwithgreenprobesbyflowcytometry.NucView™530isalsoavailableas1mMinPBS(catalognumber10408)forusewithDMSO-sensitivecelltypes.Biotiumalsooffersbluefluorogenic NucView™405Caspase-3Substrate,andgreenfluorogenicNucView™488Caspase-3Substrateandkits.Figure1.ExcitationandemissionspectraofNucView™530dyewithdsDNA. Figure2.Detectionofmitochondrialmembranepotential,caspaseactivity,andapoptosisinMCF-7cells.Cellswereeitheruntreated(A)ortreatedwithstaurosporineovernighttoinduceapoptosis(B),thenstainedwithMitoView™Blue,NucView™530Caspase-3Substrate,andCF™488AAnnexinVfor30minutesat37oCincellculturemediumwithnowash.HealthycellsshowmitochondrialstainingwithMitoViewBlue(cyan).ApoptoticcellsloseMitoView™Bluestaining(cyan),andshowNucView™520stainingofnuclei(red)andCF™488AAnnexinVstainingofcellmembranes(green). Figure3.JurkatcellsweretreatedwithDMSO(negativecontrol,blue)or0.1uM,0.25uM,or1uMstaurosporine(red)toinduceapoptosisinthepresenceofNucView™530andCF™405MAnnexinVforfourhoursat37oCbeforeanalysisonaBDLSRIIflowcytometer.StainingwithNucView™530andCF™405MAnnexinVincreasewithincreasingconcentrationofstaurosporine.CydyeisaregisteredtrademarkofGEHealthcare.

EB(溴化乙锭):溴化乙锭是一种高度灵敏的荧光染色剂,用于观察琼脂糖聚丙烯酰胺凝胶中的DNA。溴化乙锭用标准302nm 紫外光透射仪激发并放射出橙红色信号,可用Polaroid 底片或带CCD 成像头的凝胶成像处理系统拍摄。

 

SYBR Green I/II是一种结合于所有dsDNA双螺旋小沟区域的具有绿色激发波长的染料。在游离状态下,SYBR Green I发出微弱的荧光,但一旦与双链DNA结合后,荧光大大增强。因此,SYBR Green I的荧光信号强度与双链DNA的数量相关,可以根据荧光信号检测出PCR体系存在的双链DNA数量。SYBR Green I 的最大吸收波长约为497nm,发射波长最大约为520nm