4000-520-616
欢迎来到免疫在线!(蚂蚁淘生物旗下平台)  请登录 |  免费注册 |  询价篮
aatbio(优势品牌)
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
咨询热线电话
4000-520-616
当前位置: 首页 > 产品中心 > Other_kits > AAT生物任务/屏幕任务与交易;膜电位分析试剂盒*橙色荧光*/35999/1平板
商品详细AAT生物任务/屏幕任务与交易;膜电位分析试剂盒*橙色荧光*/35999/1平板
AAT生物任务/屏幕任务与交易;膜电位分析试剂盒*橙色荧光*/35999/1平板
AAT生物任务/屏幕任务与交易;膜电位分析试剂盒*橙色荧光*/35999/1平板
商品编号: 35999
品牌: aatbio
市场价: ¥56560.00
美元价: 33936.00
产地: 美国(厂家直采)
公司:
产品分类: 其它检测试剂盒
公司分类: Other_kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)535/560
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryIonChannels
MembranePotentials
RelatedBiochemicalAssays
Membranepotentialisthedifferenceinvoltagebetweentheinteriorandexteriorofacell.Themembranepotentialallowsacelltofunctionasabattery,providingpowertooperateavarietyof"moleculardevices"embeddedinthemembrane.Inelectricallyexcitablecellssuchasneurons,membranepotentialisusedfortransmittingsignalsbetweendifferentpartsofacell.Openingorclosingofionchannelsatonepointinthemembraneproducesalocalchangeinthemembranepotential,whichcauseselectriccurrenttoflowrapidlytootherpointsinthemembrane.Ionchannelshavebeenidentifiedasimportantdrugdiscoverytargets.OurScreenQuest™MembranePotentialAssayKitisahomogeneousassaywithfastreadtime.Itusesourproprietarylongwavelengthmembranepotentialindicatortodetectthemembranepotentialchangethatiscausedbytheopeningandclosingoftheionchannels.Theredfluorescenceofthemembranepotentialindicatorusedinthekithasenhancedfluorescenceuponenteringcellsandminimizestheinterferencesresultedfromthescreeningcompoundsand/orcellularautofluorescence.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.PrepareCells

 

1.1   Foradherentcells,platecellsovernightingrowthmediumat40,000to80,000cells/well/100µlfor96-wellor10,000to20,000cells/well/25µlfor384-wellplates.

1.2   Fornon-adherentcells,centrifugethecellsfromtheculturemediumandthensUSPendthecellpelletsinequalamountofHHBSandMPdye-loADIngsolution(seeSteps2.3below)at125,000to250,000cells/well/100µlfor96-wellor30,000to60,000cells/well/25µlfor384-wellpoly-Dlysineplates. Centrifugetheplatesat800rpmfor2minuteswithbreakoffpriortotheexperiments

Note: Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforthemembranepotentialchange.

 

2.PrepareMPsensordye-loadingsolution(for1plate)

 

2.1   Thaw1vialofComponentA(MPsensor),ComponentB(10XAssaybuffer)andComponentC(HHBS)atroomtemperaturebeforeuse.

Note1: 15µlofcomponentA(MPsensor)isenoughfor1plate,un-usedComponentAcanbealiquotedandstoredat<-20oCformorethan6monthsifthetubesaresealedtightly,avoidinglightandrepeatedfreeze-thawcycles.

Note2: ComponentBandCcanbestoredat4oCforconvenience.

2.2   Make1Xassaybufferbymixing1mLofComponentB(10XAssaybuffer)with9mLofComponentC(HHBS,notincludedinthekit#36001),mixthemwell.

Note: 10ml1Xassaybufferisenoughfor1plate,aliquotandstoreun-used1Xassaybufferat<-20oC,avoidlightandrepeatedfreeze-thawcycles.

 

2.3   MakeMPdye-loadingsolutionforonecellplatebyadding15µlofComponentA(MPsensor)into10mlof1Xassaybuffer(fromStep2.2),mixingthemwell. Thisworkingsolutionisstableforatleast2hoursatroomtemperature.

 

 

3. RunMembranePotentialAssay

 

3.1    Add100µL/well(96-wellplate)or25µL/well(384-wellplate)MPdye-loadingsolutionintothecellplate.

Note1: Ifyouscreencompoundsinterferewithgrowthmediumandserumfactors,thenreplacethegrowthmediumwithequalvolumeofHHBSbufferbeforeaddingtheMPdye-loadingbuffer.Alternatively,cellscanbegrowninserum-freeconditions.

Note2: DoNOTwashthecellsafterdyeloading.

3.2    Incubatethedye-loadingplateatcellincubatorfor30minutes.

Note:Insomecases,incubationatroomtemperaturefor30to60minmayworkbetter.

3.3   PreparethecompoundplatesbyusingHHBSoryourdesiredbuffer.

3.4   RunthemembranepotentialassaybymonitoringthefluorescenceatEx/Em=530/570nm.

Note:Itisimportanttorunthesignaltestbeforeyourexperiment. Differentinstrumentshavetheirownintensityrange.Adjustthesignaltestintensitytothelevelof10%to15%ofthemaximuminstrumentintensitycounts. Forexample,themaximumfluorescenceintensitycountforFLIPR-384is65,000,sotheinstrumentsettingsshouldbeadjustedtohaveitssignaltestintensityaround7,000to10,000.

References&Citations
PrinterFriendlyVersion

1.   VasilyevDV,ShanQJ,LeeYT,SolovevaV,NawoschikSP,KaftanEJ,DunlopJ,MayerSC,BowlbyMR.(2009)Anovelhigh-throughputscreeningassayforHCNchannelblockerusingmembranepotential-sensitivedyeandFLIPR.JBiomolScreen,14,1119.

2.   SollyK,CassadayJ,FelixJP,GarciaML,FerrerM,StruloviciB,KissL.(2008)MiniaturizationandHTSofaFRET-basedmembranepotentialassayforK(ir)channelinhibitors.AssayDrugDevTechnol,6,225.

3.   WeinglassAB,SwensenAM,LiuJ,SchmalhoferW,ThomasA,WilliamsB,RossL,HashizumeK,KohlerM,KaczorowskiGJ,GarciaML.(2008)Ahigh-capacitymembranepotentialFRET-basedassayforthesodium-coupledglucoseco-transporterSGLT1.AssayDrugDevTechnol,6,255.

4.   LiuCJ,PriestBT,BugianesiRM,DulskiPM,FelixJP,DickIE,BrochuRM,KnausHG,MiddletonRE,KaczorowskiGJ,SlaughterRS,GarciaML,KohlerMG.(2006)Ahigh-capacitymembranepotentialFRET-basedassayforNaV1.8channels.AssayDrugDevTechnol,4,37.

5.   BenjaminER,SkeltonJ,HanwayD,OlanrewajuS,PruthiF,IlyinVI,LaveryD,VictorySF,ValenzanoKJ.(2005)Validationofafluorescentimagingplatereadermembranepotentialassayforhigh-throughputscreeningofglycinetransportermodulators.JBiomolScreen,10,365.

6.   FelixJP,WilliamsBS,PriestBT,BrochuRM,DickIE,WarrenVA,YanL,SlaughterRS,KaczorowskiGJ,SmithMM,GarciaML.(2004)Functionalassayofvoltage-gatedsodiumchannelsusingmembranepotential-sensitivedyes.AssayDrugDevTechnol,2,260.

7.   JensenAA,Brauner-OsborneH.(2004)PharmacologicalcharacterizationofhumanexcitatoryaminoacidtransportersEAAT1,EAAT2andEAAT3inafluorescence-basedmembranepotentialassay.BiochemPharmacol,67,2115.

8.   JensenAA,KristiansenU.(2004)Functionalcharacterisationofthehumanalpha1glycinereceptorinafluorescence-basedmembranepotentialassay.BiochemPharmacol,67,1789.

9.   DavidLS,PlakasSM,ElSaidKR,JesterEL,DickeyRW,NicholsonRA.(2003)Arapidassayforthebrevetoxingroupofsodiumchannelactivatorsbasedonfluorescencemonitoringofsynaptoneurosomalmembranepotential.Toxicon,42,191.

10.   FitchRW,XiaoY,KellarKJ,DalyJW.(2003)Membranepotentialfluorescence:arapidandhighlysensitiveassayfornicotinicreceptorchannelfunction.ProcNatlAcadSciUSA,100,4909.


ViewAllAsPDF
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
联络我们
服务热线:4000-520-616
(限工作日9:00-18:00)
QQ :1570468124
手机:18915418616