msd-sulfo-tag-nhs-ester-说明书

18153-v1-2015Feb|1MSDGOLD™SULFO-TAGNHS-EsterMSDGOLDSULFO-TAG™NHS-Ester150nmol(sufficientforconjugating1mgofIgG)R91AO-12µmol(sufficientforconjugating10mgofIgG)R91AO-2MSDGOLDSULFO-TAGNHS-EsterConjugationPacksPack1(sufficientforconjugating5x200µgofIgG)R31AA-1Pack2(sufficientforconjugating5x1mgofIgG)R31AA-218153-v1-2015Feb|2MSDLabelingReagentsMSDGOLDSULFO-TAGNHS-EsterForlabelingaminesNote:150nmolsizeofMSDGOLDSULFO-TAGNHS-Esterissufficientforconjugating1mgofIgGandthe2µmolsizeissufficientforconjugating10mgofIgGatachallengeratioof20.EachMSDGOLDSULFO-TAGNHS-EsterConjugationPackcontainsenoughmaterialfor5reactions.Atachallengeratioof20,Pack1issufficientforconjugating200µgofIgGperreaction,andPack2issufficientforconjugating1mgofIgGperreaction.Thispackageinsertmustbereadinitsentiretybeforeusingthisproduct.FORRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.MESOSCALEDISCOVERY®AdivisionofMesoScaleDiagnostics,LLC.1601ResearchBoulevardRockville,MD20850-3173USA(design),96WELL1,4,7,&10-SPOT(designs),384WELL1&4-SPOT(designs),MSD(design),U-PLEX(design),V-PLEX(design),S-PLEX(design),andSPOTTHEDIFFERENCEaretrademarksand/orservicemarksofMesoScaleDiagnostics,LLC.©2015MesoScaleDiagnostics,LLC.Allrightsreserved18153-v1-2015Feb|3TableofContentsIntroduction..................................................................................................................................................................4PreparationofMSDGOLDSULFO-TAGConjugates........................................................................................................5MSDGOLDSULFO-TAGNHS-EsterConjugationPacks..................................................................................................6AdditionalMaterialsandEquipment...............................................................................................................................6Protocol.......................................................................................................................................................................8Storage,Handling,andStability..................................................................................................................................10FAQs..........................................................................................................................................................................11Worksheet..................................................................................................................................................................14OrderingInformationMSDCustomerServicePhone:1-240-314-2795Fax:[phone]Email:[e-mail]hone:1-240-314-2798Fax:[phone]attn:ScientificSupportEmail:[e-mail]18153-v1-2015Feb|4IntroductionThisprotocoldetailstheconjugationprocedureforproteinsofmolecularweight(MW)40,000DaltonsusingMSDGOLDSULFO-TAGNHS-Esterlabel.Thestraightforwardprocedureinvolvesanoptionalbufferexchangestep,a2-hourincubationstep,andamandatorybufferexchangesteptoquicklyisolatetheconjugatedproteinusingaspincolumn.MSDGOLDSULFO-TAGNHS-Ester(Figure1)isanaminereactive,N-hydroxysuccinimideesterwhichreadilycouplestoprimaryaminegroupsofproteinsundermildlybasicconditionstoformastableamidebond.MSDGOLDSULFO-TAGconjugatesarestableandmaybeusedatlowconcentrations.Thesefeaturesminimizetime,cost,andlaboraslargebatchesofastableconjugatecanbeprepared,validated,andusedforlongperiodsoftime.ItsexcellentperformancecharacteristicsandsimpleconjugationproceduremakeMSDGOLDSULFO-TAGNHS-Estertheproductofchoiceformoleculesthatcontainprimaryamines(e.g.,lysine-containingproteins).MSDGOLDSULFO-TAGofferslownon-specificbinding,resultinginhighlysensitivedetectionwhenusedinconjunctionwithMSDinstruments.Figure1:MSDGOLDSULFO-TAGNHS-Ester18153-v1-2015Feb|5PreparationofMSDGOLDSULFO-TAGConjugatesGeneralNotesInordertominimizehydrolysisofMSDGOLDSULFO-TAGNHS-Ester,thereagentshouldbedissolvedincolddistilledwaterjustpriortoitsadditiontotheproteinsolution.Ifnecessary,thestockMSDGOLDSULFO-TAGNHS-Estersolutioncanbekeptoniceforupto10minutes.Thereconstitutedsolutionisunstableandanyunusedmaterialshouldbediscarded.ConsiderconjugatingmorethanoneproteinatthesametimetomaximizetheuseoftheMSDGOLDSULFO-TAGNHS-Esterreagent.Generally,150nmolofMSDGOLDSULFO-TAGNHS-Esterissufficientforconjugationofupto1mgofIgG.ThelabelingratioisthenumberofmoleculesofMSDGOLDSULFO-TAGconjugatedtoeachmoleculeofprotein.OptimalconjugationratiosforaMSDGOLDSULFO-TAGconjugatedproteinshouldbedeterminedempiricallyforeachspecificapplication.FormostapplicationsusingIgGantibodies(MW~150,000),optimalperformanceisobtainedwithconjugationratiosbetween2:1and20:1.Inthisrange,assaysignalstypicallyshowalineardependenceonconjugationratio.Conjugationratiossignifica