| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 346/617 |
| MW | 1116.96 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | SuperiorLabelingDyes trFluor™DyesandKits |
| Related | LabelingviaThiolGroups |
| Spectrum | AdvancedSpectrumViewer |
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 |  PrinterFriendlyVersion |
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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.
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