TUBEs: ARevolutioninPolyubiquitinIsolationandCharacterization
TheuseofTandemUbiquitinBindingEntities(TUBEs)isemergingasanindispensablestrategyforubiquitinresearch.ThefirstgenerationoftheseTUBEsbindK48-andK63-linkedtetraUbchainswithsingledigitnanomolarKds,~100to1000-foldmoretightlythanmonomericUBAs.TUBEsalsoprotectproteinsfromDUBsandtheproteasome,evenintheabsenceofinhibitorsnormallyrequiredtoblocksuchactivity.ThisallowsefficientisolationofnativepolyUbchainsandattachedproteinsfromcelllines,tissues,andorgansunderconditionsthatarelesslikelytoaltercellphysiologythanthoselistedabove.TUBE1andTUBE2havebeendemonstratedtoenrichforallpolyUbchainlinkagetypes,withoutdiscrimination,makingthesereagentsappropriateevenifthelinkagetypeisnotknownfortheproteinofinterest.
ThenextgenerationofTUBEs: LinkageSpecificIsolationofPolyUbChains
K48TUBEHFwasdevelopedtoshowenhancedselectivityforK48-linkedpolyubiquitinchains(~20nM)overallotherlinkages(>2µM).ItcanbeusedaloneorinconjunctionwithourotherTUBEproducts,especiallyK63TUBEandM1(linear)TUBEtoinvestigatepolyubiquitinchainlinkageinyoursubstrateprotein.
Note:ThenewK48TUBEHF(UM607)hasacomparableaffinityasourexistingK63TUBE(UM604).BothUM607andUM604bindtotheircognateUb4swithaKd of~10-20nM.PleasenotethattheoldK48TUBE(UM605)bindsK48-Ub4 withaKd of~200nM.
Info
Tag: FLAG
Size:50µg
Buffer:PBS,pH7.2
Storage: Stableforupto6monthsat-80C.Aliquotreagentandavoidrepeatedfreeze/thawcycles.
APPLICATIONS
- Far-WesterndetectionofK48-polyubiquitinatedproteinsfromcellandtissueextracts
- IsolationandenrichmentofK48-polyubiquitinatedproteinsfromcellandtissueextracts
- IsolateK48-polyubiquitinatedproteinsforproteomicstudies
BENEFITS
- NanomolaraffinityforK48poly-ubiquitinchains
- 100-foldpreferenceforK48poly-ubiquitinchainsoverallotherlinkages
- TUBEsdisplayhigheraffinitytowardspolyubiquitinsthanmostubiquitinantibodies
- Avoidsoverexpressionofepitope-taggedubiquitinforpulldowns
- CompatIBLewithFLAG® technology,providingflexibilityandspecificity
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