Only1hourtorecoverconjugates

AllprocessesinasingleFiltrationtube

Highrecoveryofconjugates

Applicablefor50-200μgIgG

FluoresceinLabellingKit-NH2ismainlyusedforthepreparationoffluorescein-labelledproteinssuchasIgGforimmunostainingandcellularproteinsfortracing.Amine-reactivefluorescein,acomponentofthiskit,hassuccinimidylgroups(NHS)thatreactwiththeaminogroupsonproteinsorothermolecules.

Thiskitcontainsallthereagentsnecessaryforlabelling,includingstoragebuffer.Eachvialoffluoresceincanlabelupto200μgofIgG,conjugatingabout4to6fluoresceinmoleculesperIgGmolecule.Becausethiskitalsoincludesabufferexchangesystem,asamplecontainingaminebasebuffercanbelabelled.AlthoughmembranefiltrationsometimescausesIgGaggregation,thebuffersysteminthiskitpreventsaggregationduringtheconcentrationofIgGorfluorescein-labelledIgGsolution.Afluorescein-labelledIgGsolutionpreparedusingthiskitisstableformorethan2monthsat4ºC.TheexcitationandemissionwavelengthsofthefluoresceinlabelledIgGare495nmand520nm,respectively.

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Q&A

CanIusethiskitforotherproteins?
Yes,ifthemolecularweightisgreaterthan50,000.

DoIhavetouseaFiltrationtubepriortolabelingtheprotein?<
Iftheproteinsolutiondoesnotcontainsmallmoleculeswithanaminogroupandtheconcentrationoftheproteinis10mgperml,orabout70μM,thereisnoneedtousetheFiltrationtube.Justmix10μlofthesamplesolutionwith90μlofReactionbufferandadd8μlNH2-reactivefluorescein(preparedatstep3)tothemixture,andfollowtheprotocolstartingatstep4.

Howlongisthefluorescein-labeledproteinstable?
Ifyoustoreat4ºC,itisstableforover2months.Forlongerstorage,add100%volumeofglycerol,aliquot,andstoreat-20ºC.However,pleasenotethatstABIlitydependsontheproteinitself.<

WhatistheminimumamountofIgGthatcanbelabeledbythiskit?
TheminumumamountofIgGis10μg;simplyfollowtheprotocol.Thelabelingratioremainsthesamefor10μgto100μgofIgG.

CanIusethiskittolabeloligonucleotidesorpeptides?
No.OligonucleotidesandpeptidesmaybetoosmalltoretainonthemembranefilteroftheFiltrationtube.