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主营:主营:研究并生产荧光和发光探针,信号转导研究的试剂
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当前位置: 首页 > 产品中心 > Fluorescent_dyes > AAT Bioquest/细胞仪与贸易;凋亡和坏死复合检测试剂盒I*三重荧光颜色*/22840/100 Te
商品详细AAT Bioquest/细胞仪与贸易;凋亡和坏死复合检测试剂盒I*三重荧光颜色*/22840/100 Te
AAT Bioquest/细胞仪与贸易;凋亡和坏死复合检测试剂盒I*三重荧光颜色*/22840/100 Te
AAT Bioquest/细胞仪与贸易;凋亡和坏死复合检测试剂盒I*三重荧光颜色*/22840/100 Te
商品编号: 22840
品牌: aatbio
市场价: ¥71060.00
美元价: 42636.00
产地: 美国(厂家直采)
公司:
产品分类: 荧光染料
公司分类: Fluorescent_dyes
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
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Ex/Em(nm)None/None
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryCellAnalysis
CellCytotoxicity
RelatedApoptosisandCytotoxicity
CellApoptosis
BiochemicalAssays
OurCellMeter™assaykitsareasetoftoolsformonitoringcellviABIlity.Thereareavarietyofparametersthatcanbeused.Thisparticularkitisdesignedtomonitorcellapoptotic,necroticandhealthycells.Apoptosisisdescribedasanactive,programmedprocessofautonomouscellulardismantlingthatavoidselicitinginflammation.Inapoptosis,phosphatidylserine(PS)istransferredtotheouterleafletoftheplasmamembrane.Asauniversalindicatoroftheinitial/intermediatestagesofcellapoptosis,theappearanceofphosphatidylserineonthecellsurfacecanbedetectedbeforemorphologicalchangesareobserved.ThePSsensorusedinthiskithasgreenfluorescence(Ex/Em=490/525nm)uponbindingtomembranePS.Necrosishasbeencharacterizedaspassive,accidentalcelldeathresultingfromenvironmentalperturbationswithuncontrolledreleaseofinflammatorycellularcontents.Lossofplasmamembraneintegrity,asdemonstratedbytheabilityofamembrane-impermeable7-AAD(Ex/Em=546/647nm)tolabelthenucleus,representsastraightforwardapproachtodemonstratelatestageapoptosisandnecrosis.Inaddition,thiskitalsoprovidesalivecellcytoplasmlabelingdyeCytoCalcein™Violet450(Ex/Em=405/450nm)forlabelinglivingcellcytoplasm.Thiskitisoptimizedtodetectcellapoptosis(green),necrosis(greenand/orred)andhealthycells(blue)withaflowcytometerandfluorescencemicroscope.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

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


品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。 1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物; 2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞; 3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类; 4)致力于开发用于信号转导研究的试剂; 5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
品牌分类
淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
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