Description

Whenyouwishtocovalentlycoupleantibodies,peptides,intactproteinsandfunctionalenzymestothesurfaceofamagneticbeadchooseDynabeads®Tosylactivated.Optimalantibodyorientationmakesthesebeadsanexcellentchoicefortheimmunoprecipitationofproteinsandproteincomplexes.

IdealforImmunoprecipitationofproteincomplexes:
LowbackgroundandcovalentlinkingofantibodiestothebeadsurfacemakeDynabeadsTosylactivatedanexcellentchoiceforImmunoprecipitationofproteinsandproteincomplexes(Co-Immunoprecipitation,Co-IP).Gentleandrapidmagneticconcentrationofthebeadsandshortincubationtimes(possIBLeduetofastsurface-basedbindingkinetics)makeDynabeadsTosylactivatedanexcellentchoicefortheimmunoprecipitationofhighly-lABIleorand/ortransient(short-lived)proteincomplexes.

Ultrarapidprotocols:

•Identifymembersofproteincomplexesinminutesratherthanhours
•Temporalresolutionshortenoughtoidentifytransientandlabilecomplexes
•Identifybindingpartnersthatcannotbeidentifiedwithlongerprotocols
•Rapidprotocolsfurtherreducethealreadyultra-lowbackgroundbinding
•Strongersignaltonoise-ratios

Applications:
•IPofproteinsandproteincomplexes
•Couplefunctionalenzymestobeadsurfacefordownstreamassays
•Couplepeptidestobeadsurfacetoidentifybinders

Purifypeptides,proteinsandenzymesthatare:
•Transientlystable
•Structurallyintact
•Temperaturelabile
•Intheirnativeconformationandfunctional

CouplingProcedureOutline:
CovalentcouplingisperformedovernightbyincubatingthedesiredligandwiththeDynabeadsTosylactivated.LigandscommonlycoupledtoDynabeadsTosylactivatedincludepeptidesandproteins(e.g.antibodiesforImmunoprecipitationorCo-Immunoprecipitation).CouplingoccursatneutraltohighpHandat37°C.WerecommendcouplingatpH8.5-9.5,butforpHlabileligands,couplingcanbeperformedinanalternativebufferatpH7.4.

Uponcompletionoftheligandcouplingstep,theactualDynabeadsTosylactivatedsurfacecoatingwillberenderedinert,resultinginlownon-specificbinding.

AboutDynabeads:
Dynabeadsarenon-porousmonodispersesuperparamagneticbeads.Theyarehighlymobileinsolutionenablingligandscoupledtothebeadstocontinuouslyinteractwiththeentiresamplevolume.Thesuperparamagneticbeadsarepulledtothetubewallsbytransferringthetubetoarackcontainingastrongmagneticfield(FromtherednavigationbarnavigateBrands>Dynal®>Magnets).Strongmagneticfieldsquicklypullthebeadstothetubewall,allowingforeasyandcompleteremovalofthesupernatantbyPipette.Washingstepsareperformedsimilarly.l

BeadSurfaceCharacteristicsforDynabeadsTosylactivated:
•p-toluene-sulfonyl(Tosyl)groups
•Hydrophobic,pHNeutral
•Covalentbindingbyprimaryamine(NH₂)orsulphydryl(SH)groups

Bindingcapacitypermilligrambeads:
Variesdependingonligand,(e.g.5-10µgIgG)