Thiskitissufficientforpurificationoffreepolyubiquitinchainsin100mgor20X5mgcellortissueextractsataconcentrationof5mg/ml.
DescriptionTheUnconjugated(Free)PolyubiquitinChainCaptureKityenrichfreepolyUbchainsincellortissueextractswithin6hours.ThisapproachusestheN-terminalZnF-UBPdomainofthedeubiquitinatingenzymeUsp5tospecificallycapturefreepolyUbchains.TheZnF-UBPdomainofUsp5bindstheC-terminaldiglycinemotifinfreeUbchains,itdoesnotbindpolyUbchainsconjugatedonsubstrateproteins.
Components•GST-ZnF-UBP(4mg/ml)..............................................0.5ml•GST(4mg/ml).............................................................0.5ml
•Iodoacetamine(IAA)....................................................25mg•Glutathione(reduced).................................................100mg
•GlutathioneAgaroseResin(75%slurry).....................1.0ml
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10µlGST-boundorGST-ZnF-UBP-boundproteinswereseparatedonSDS-PAGEsandimmunoblottedwithantibodiesagainUborK48-linkedpolyUbchains.