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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/trFluor™ Eu maleimide/1434/100 ug
产品编号:1434
市  场 价:¥85560.00
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:$4278.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/trFluor™ Eu maleimide/1434/100 ug
商品介绍
Overview
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Ex/Em(nm)346/617
MW1116.96
CAS#N/A
SolventDMSO
StorageF/D/L
CategorySuperiorLabelingDyes
trFluor™DyesandKits
RelatedLabelingviaThiolGroups
ManyBIOLOGicalcompoundspresentincells,serumorotherbiologicalfluidsarenaturallyfluorescent,andthustheuseofconventional,promptfluorophoresleadstoseriouslimitationsinassaysensitivityduetothehighbackgroundcausedbytheautofluorescenceofthebiologicalmoleculestobeassayed.Theuseoflong-livedfluorophorescombinedwithtime-resolveddetection(adelaybetweenexcitationandemissiondetection)minimizespromptfluorescenceinterferences.OurtrFluor™Euprobesenabletime-resolvedfluorometry(TRF)fortheassaysthatrequirehighsensitivity.ThesetrFluor™EuprobeshavelargeStokesshiftsandextremelylongemissionhalf-liveswhencomparedtomoretrADItionalfluorophoressuchasAlexaFluororcyaninedyes.ComparedtotheotherTRFcompounds,ourtrFluor™EuprobeshaverelativelyhighstABIlity,highemissionyieldandabilitytobelinkedtobiomolecules.Moreover,ourtrFluor™Euprobesareinsensitivetofluorescencequenchingwhenconjugatedtobiologicalpolymerssuchasantibodies.
SpectrumAdvancedSpectrumViewer

Sorry,yourbrowserdoesnotsupportinlineSVG.RelativeIntensity(%)100806040200Sorry,yourbrowserdoesnotsupportinlineSVG.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

SampleProtocolforLabelingProteinswithtrFluor™EuMaleimide:

1)                  Dissolveyourthiol-containingproteinatconcentration1-10mg/mL(3-10mgistheoptimallabelingconcentration)usingPBSbuffer(20mM,pH7.2).

2)                  DissolvethetrFluor™MaleimideinDMSOattheconcentrationof10-15mg/mL(Forexample,tothe100µgtrFluor™Maleimideadd10µLDMSOtomake7mMdyelabelingsolution).

3)                  MixthetrFluor™Maleimide(fromStep2)andproteinsolution(fromStep1)at20:1molarratioofdye/protein,andshakethereactionmixtureatroomtemperaturefor2-4hoursinthedark.

4)                  Filterthereactionmixturethroughaproteinspincolumnfor100µgto1mgproteinlabelingreaction;orpurifytheconjugateusinggelfiltrationonaproperlysizedSephadexG-25columnifthereactionscaleislargerthan1mg.

5)                  Collectthedesiredfractionsforyourimmediateuseorfreezedrythemforyourfutureuse.

 

Note:ThetrFluor™conjugateneedbeusednearneutralpHrange(6.5to7.5).EitheracidicorbasicpHwouldreduceitsfluorescenceintensity.

 

SampleProtocolforLabelingSmallMoleculeswithtrFluor™EuMaleimide:

1)                  DissolvetrFluor™Maleimide(10-15mg/mL)andyourthiol-containmoleculeinDMSOat1:1.2molarratioofdye/thiol-containmolecule(Forexample,tothe100µgtrFluor™Maleimideadd10µLDMSOtomake7mMdyelabelingsolution).

2)                  Stirthereactionmixtureatroomtemperaturefor2-4hoursinthedark.

3)                  PurifytheconjugateusingHPLC(ammoniumacetate/waterandacetonitrile,pH7.0).

4)                  Collectandpoolthedesiredfractions.

5)                  Combineandfreeze-drythepooledfractions.

 

Note:ThetrFluor™conjugateneedbeusednearneutralpHrange(6.5to7.5).EitheracidicorbasicpHwouldreduceitsfluorescenceintensity.

References&Citations
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1.   SavilleL,SpaisC,MasonJL,AlbomMS,MurthyS,MeyerSL,AtorMA,AngelesTS,HustenJ.(2012)Time-ResolvedFluorescenceResonanceEnergyTransferasaVersatileToolintheDevelopmentofHomogeneousCellularKinaseAssays.AssayDrugDevTechnol.

2.   LoMC,NgoR,DaiK,LiC,LiangL,LeeJ,EmkeyR,EksterowiczJ,VenturaM,YoungSW,XiaoSH.(2012)Developmentofatime-resolvedfluorescenceresonanceenergytransferassayforcyclin-dependentkinase4andidentificationofitsATP-noncompetitiveinhibitors.AnalBiochem,421,368.

3.   PailaYD,KombrabailM,KrishnamoorthyG,ChattopadhyayA.(2011)OligomerizationoftheSEROtonin(1A)receptorinlivecells:atime-resolvedfluorescenceanisotropyapproach.JPhysChemB,115,11439.

4.   MartikkalaE,Rozwandowicz-JansenA,HanninenP,Petaja-RepoU,HarmaH.(2011)Ahomogeneoussingle-labeltime-resolvedfluorescencecAMPassay.JBiomolScreen,16,356.

5.   GaboritN,LarbouretC,VallagheJ,PeyrussonF,Bascoul-MolleviC,CrapezE,AzriaD,ChardesT,PoulMA,MathisG,BazinH,PelegrinA.(2011)Time-resolvedfluorescenceresonanceenergytransfer(TR-FRET)toanalyzethedisruptionofEGFR/HER2dimers:anewmethodtoevaluatetheefficiencyoftargetedtherapyusingmonoclonalantibodies.JBiolChem,286,11337.

6.   LeyrisJP,RouxT,TrinquetE,VerdieP,FehrentzJA,OueslatiN,DouzonS,BourrierE,LamarqueL,GagneD,GalleyrandJC,M"KadmiC,MartinezJ,MaryS,BaneresJL,MarieJ.(2011)Homogeneoustime-resolvedfluorescence-basedassaytoscreenforligandstargetingthegrowthhormonesecretagoguereceptortype1a.AnalBiochem,408,253.

7.   PosokhovYO,KyrychenkoA,LadokhinAS.(2010)Steady-stateandtime-resolvedfluorescencequenchingwithtransitionmetalionsasshort-distanceprobesforproteinconformation.AnalBiochem,407,284.

8.   Alvarez-CurtoE,WardRJ,PedianiJD,MilliganG.(2010)LigandregulationofthequaternaryorganizationofcellsurfaceM3muscarinicacetylcholinereceptorsanalyzedbyfluorescenceresonanceenergytransfer(FRET)imagingandhomogeneoustime-resolvedFRET.JBiolChem,285,23318.

9.   KotaS,ScampaviaL,SpicerT,BeelerAB,TakahashiV,SnyderJK,PorcoJA,HodderP,StrosbergAD.(2010)Atime-resolvedfluorescence-resonanceenergytransferassayforidentifyinginhibitorsofhepatitisCviruscoredimerization.AssayDrugDevTechnol,8,96.

10.   VisserAJ,LaptenokSP,VisserNV,vanHoekA,BirchDJ,BrochonJC,BorstJW.(2010)Time-resolvedFRETfluorescencespectroscopyofvisiblefluorescentproteinpairs.EurBiophysJ,39,241.


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