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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Cell Meter™ Multiplexing Live, Apoptotic and Necrotic Cell Detection Kit III *Triple Fluorescence Colors*
产品编号:22846
市  场 价:¥143560.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$7178.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Cell Meter™ Multiplexing Live, Apoptotic and Necrotic Cell Detection Kit III *Triple Fluorescence Colors*
商品介绍
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Ex/Em(nm)None/None
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryCellAnalysis
CellCytotoxicity
RelatedApoptosisandCytotoxicity
CellApoptosis
OurCellMeter™assaykitsareasetoftoolsformonitoringcellviABIlity.Thereareavarietyofparametersthatcanbeused.Thisparticularkitisdesignedtosimultaneouslymonitorapoptotic,necroticandhealthycells.Apoptosisisanactive,programmedprocessofautonomouscellulardismantlingthatavoidselicitinginflammation.Inapoptosis,phosphatidylserine(PS)istransferredtotheouterleafletoftheplasmamembrane.Asauniversalindicatoroftheinitial/intermediatestagesofcellapoptosis,theappearanceofphosphatidylserineonthecellsurfacecanbedetectedbeforemorphologicalchangesareobserved.ThePSsensorAnnexinV-iFluor™647conjugatehasredfluorescence(Ex/Em=620/660nm)uponbindingtomembranePS.Necrosishasbeencharacterizedaspassive,accidentalcelldeathresultingfromenvironmentalperturbationswithuncontrolledreleaseofinflammatorycellularcontents.Lossofplasmamembraneintegrity,asdemonstratedbytheabilityofamembrane-impermeableNuclearBlue™DCS1(Ex/Em=360/450nm)tolabelthenucleus,representsastraightforwardapproachtodemonstratelatestageofapoptosisandnecrosis.Inaddition,thiskitalsoprovidesalivecelllabelingdye,Cellbrite™Orange(Ex/Em=550/570nm),forlabelingnon-apoptotichealthycells.Thiskitisoptimizedtosimultaneouslydetectcellapoptosis(red),necrosis(blueand/orred)andhealthycells(orange)withafluorescencemicroscope.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. Preparecells:
    Plate100to100,000cells/wellinatissueculturemicroplatewithblackwallandclearbottom.Addtestcompoundsintothecellsandincubateforadesiredperiodoftime(suchas24,48or96hours)ina37°C,5%CO2incubator.Forblankwells(mediumwithoutthecells),addthesameamountofcompoundbuffer.Thesuggestedtotalvolumeis100µL/well/96-wellplateand25µL/well/384-wellplate.
    Note:WetreatedHeLacellswithstaurosporine(SS)for4hoursat37ºCtoinducecellapoptosis.SeeFigure1fordetails.

  2. Preparetriplefluorescenceassaysolution:
    1. Preparetriplefluorescenceassaysolutionbyadding10μLof100XAnnexinV-iFluor™488conjugate(ComponentA),5μLof200XNuclearBlue™DCS1(ComponentC)and5μLof200XCellbrite™Red(ComponentD)to1mLofAssayBuffer(ComponentB).Thetriplefluorescenceassaysolutionisstableforatleast1houratroomtemperature.
      Note:Astheoptimalstainingconditionsmayvarydependingondifferentcelltypes,it’srecommendedtodeterminetheappropriateconcentrationofComponentA,CandDindividually.
    2. Removecellculturemediumandtestcompoundsaftertreatment(fromStep1).
    3. Add100µL/well(96-wellplate)or25µL/well(384-wellplate)oftriplefluorescenceassaysolution.Incubateatroomtemperatureor37ºCfor30to60minutes,protectedfromlight.
    4. WashcellswithHBSS,PBSorbufferofyourchoicetwice.
    5. AnalyzetheapoptoticcellswithAnnexinV-iFluor™488underfluorescencemicroscopewithaFITCfilter.ThegreenstainingontheplasmamembraneindicatestheAnnexinV-iFluor™488conjugatebindingtoPSoncellsurface.MonitorthefluorescenceintensitywithaDAPIfilterfornecrosis,TexasRedorCy5filterforlivecellsusingafluorescencemicroscope(SeeFigure1fordetails).
References&Citations
CitationExplorer

Anthocyanin-richblackcurrantextractinhibitsproliferationoftheMCF10AhealthyhumanbreastepithelialcelllinethroughinductionofG0/G1arrestandapoptosis
Authors:NaokiNanashima,KayoHorie,MitsuruChiba,ManabuNakano,HayatoMaeda,ToshiyaNakamura
Journal:MolecularMedicineReports(2017):6134--6141

ClusterinsignalsviaApoER2/VLDLRandinducesmeiosisofmalegermcells
Authors:MuhammadAssadRiaz,AngelikaStammler,MareikeBorgers,LutzKonrad
Journal:AmericanJournalofTranslationalResearch(2017):1266

DetectingApoptosis,Autophagy,andNecrosis
Authors:JackColeman,RuiLiu,KathyWang,ArunKumar
Journal:ApoptosisMethodsinToxicology(2016):77--92


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