Click-&-GoTMPlusProteinEnrichmentKitisanefficient,biotin-streptavidinfreetoolforcaptureofalkyne-modifiedproteinsonapicolylazide-agaroseresin.Picolylazidemoietyisanuniqueazidethatincorporatesacopper-chelatingmotiftoraisetheeffectiveconcentrationofCu(I)atthereactionsite.TheincorporationofcopperchelatingmoietyintoazidereportsisthelateststepinimprovingkineticsofCuAAC,whichinturnleadstoasubstantialincreaseinthesensitivityofalkynedetection.Thisisofspecialvalueforthedetectionoflowabundancetargetsorwhereseveralincreaseofsignalintensityisdesired.
Learnmoreaboutpicolylazidereagents.
Thealkynemodificationcanoccurviametabolicfeeding,enzymaticaddition,orchemicalmodification.Alkyne-modifiedproteins,ortheirpost-translationallymodifiedforms,arecapturedfromcomplexproteinextractsontheazide-agaroseresinsupplied.OncecovalentlyattachedtotheresinviacoppercatalyzedClickchemistry,beadscanbewashedwithhigheststringencyvirtuallyeliminatinganynon-specificallyboundproteinstoyieldahighlyenrichedpopulationofnascentmolecules.Uponproteasedigestion,thisyieldsahighlypurepeptidepoolthatisidealformassspectrometry(e.g.,LCMS/MS)basedanalysis.
Thebiotin-freeenrichmentimprovessignaltonoisebyeliminatingnonspecificbindingandincreasingselectivity,thusimprovingdetectionoflowabundantproteins.ThisapproachisfullycompatIBLewithwidelyusedMStechniquesincludingiTRACandICAT.
Thebiotin-free,covalentcaptureapproachissuperiortobiotinorlectinbasedenrichmentapproaches,andidealforproteomics,bioMarkerdiscovery,posttranslationalmodification(PTM)analysis,andmore.
TheClick-&-GoTMPlusProteinEnrichmentKitcanbeusedasdirectreplacementofClick-iT®ProteinEnrichmentKit
| Enrichmenttarget | Alkyne-taggedproteins |
| Format | SpinColumn |
| Isolationtechnology | Biotin-freecovalentcapture |
| StorageCondition | 4C |
| Shippingconditions | Ambienttemperature |
