| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | None/None |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellAnalysis CellCytotoxicity |
| Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
1.PrepareandincubatecellswithApopxin™Green:
1.1 Treatcellswithtestcompoundsforadesiredperiodoftime(4-6hoursforJurkatcellstreatedwithstaurosporine)toinduceapoptosis.
1.2 Centrifugethecellstoget1-5×105cells/tube.
1.3 ResUSPendcellsin200μLofAssayBuffer(ComponentB).
1.4 Add2μLofApopxin™Green(ComponentA)intothecells.
Optional1:Add1µLof200X7-AAD(ComponentC)intothecellsfornecrosiscells.
Optional2:Add100µLofDMSOintothevialofCytoCalcein™Violet450(ComponentD)tohave200XCytoCalcein™Violet450stocksolution,andthenadd1µLintothecellsforhealthycellsstaining.
1.5 Incubateatroomtemperaturefor30to60minutes(protectedfromlight).
1.6 Add300μLofAssayBuffer(ComponentB)toincreasevolumebeforeanalyzingthecellswithaflowcytometerorfluorescencemicroscope(seeStep1.7below).
1.7 MonitorthefluorescenceintensityatEx/Em=490/525nmforapoptosis,550/650nmfornecrosis,and405/450nmforhealthycellsusingaflowcytometerorafluorescencemicroscope(SeeStep2or3below).
2.Analyzecellsusingaflowcytometer:
QuantifyApopxin™GreenbindingbyusingtheFL1channel(Ex/Em=490/525nm),andmeasurethecellviabilityusingtheFL3channel(Ex/Em=490/650nm)when7-AADisadded,and/orusingEx/Em=405/450nmwhenCytoCalcein™Violet450isaddedintothecells.
Note:TheflowcytometricanalysisofApopxin™bindingtoadherentcellsisnotroutinelytestedsincespecificmembranedamagemayoccurduringcelldetachmentorharvesting.However,methodsforutilizingAnnexinVforflowcytometryonadherentcelltypeshavebeenpreviouslyreportedbyCasiola-Rosenetal.andvanEngelendetal(seeRefs1and2).
3.Analyzecellsusingafluorescencemicroscope:
3.1 PipettethecellsuspensionfromStep1.5,rinse1-2timeswithassaybuffer,andthenresuspendthecellswithassaybuffer.Addthecellsonaglassslidethatiscoveredwithaglasscoversliporablackwall/clearbottom96-wellmicroplate.
Note:Foradherentcells,itisrecommendedtogrowthecellsdirectlyonacoverslip(orablackwall/clearbottom96-wellmicroplate).AfterincubationwithApopxin™Green(Step1.5),rinse1-2timeswithassaybuffer,andthenaddassaybufferbacktothecoverslip(orablackwall/clearbottom96-wellmicroplate).Invertcoversliponaglassslideandvisualizethecells.Thecellscanalsobefixedin2%formaldehydeaftertheincubationwithApopxin™Greenandvisualizedunderamicroscope.
3.2 AnalyzetheapoptoticcellswithApopxin™GreenunderafluorescencemicroscopeusingtheFITCchannel.MeasurethecellviabilityusingtheTexasRedchannelwhen7-AADisadded,and/orVioletchannelwhenCytoCalcein™Violet450isaddedintothecells.ThegreenstainingontheplasmamembraneindicatestheApopxin™GreenbindingtoPSoncellsurface.
| 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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然后是碱性荧光染料，造纸厂用来增加纸的亮度。
直接和分散的荧光黄，印染厂用来增加棉质和化纤面料的艳度
For principle,look at this site:
碘化丙啶(propiolium iodide,PI)能嵌入DNA双螺旋中,可使荧光强度增加约20倍,以488nm波长激发,DNA/PI复合物最大的发射波长约为615nm.
1.小鼠Lewis肺癌细胞DNA含量测定方法
(1).从C57BL/6小鼠上切除肿块,在培养皿内用PBS冲洗.
(2).去除结缔组织及脂肪,剪碎肿块.
(3).小碎片移入1.20×38mm注射针,加压使其通过,于4℃条件下重悬细胞于HBSS中.
(4).将200～300μL细胞悬液(5×105细胞/mL)中加入3mL PI(50μg/mL),染色3LL细胞,于4℃存放20～30分钟.
(5).测定580～750nm之间的发射荧光,以去除末结合PI产生的激发光与发射光谱线之间的重叠部分.
是欢快活泼的光辉色彩，是暖色系中最温暖的色，它使人联想到金色的秋天，丰硕的果实；
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橙色也会带来一种甜甜的感觉。
