The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Typical enzyme concentration to support hydrolysis of substrates in vitro ranges from 10-100 nM depending on conditions and substrate. Pre-incubation (15 min) with 10 mM DTT is recommended to achieve maximum activity.
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Preparation and Storage
Stability and Storage
Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.
Ubiquitin specific protease 5 (ubiquitin isopeptidase T)
Ubiquitin thioesterase 5
Ubiquitin thiolesterase 5
Ubiquitin-specific-processing protease 5
Cleaves linear and branched multiubiquitin polymers with a marked preference for branched polymers. Involved in unanchored 'Lys-48'-linked polyubiquitin disassembly. Binds linear and 'Lys-63'-linked polyubiquitin with a lower affinity. Knock-down of USP5 causes the accumulation of p53/TP53 and an increase in p53/TP53 transcriptional activity because the unanchored polyubiquitin that accumulates is able to compete with ubiquitinated p53/TP53 but not with MDM2 for proteasomal recognition.
Belongs to the peptidase C19 family. Contains 2 UBA domains. Contains 1 UBP-type zinc finger.
The UBP-type zinc finger domain interacts selectively with an unmodified C-terminus of the proximal ubiquitin. Both UBA domains are involved in polyubiquitin recognition.