详细说明
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.
Activity
Recombinant Human USP12/UAF1 Complex is a Ubiquitin-specific deconjugating enzyme heterodimer. Reaction conditions will need to be optimized for each specific application. We recommend an initial USP12/UAF1 Complex concentration of 50-500 nM when using Ubiquitin-AMC or Ubiquitin-Rhodamine (U-550, U-555) as a substrate. In vitro, the USP12/UAF1 complex will not efficiently hydrolyze poly-Ubiquitin chains in the absence of WDR20.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived Contains a C-terminal 6-His tag
Accession #
Predicted Molecular Mass
44 kDa (USP12), 77 kDa (UAF1)
E-580 |
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Formulation X mg/ml (X μM) in 50 mM HEPES pH 8.0, 150 mM NaCl, 10% (v/v) Glycerol, 2 mM TCEP | ||
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. | ||
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Background: USP12/UAF1 Complex
Ubiquitin carboxyl-terminal hydrolase 12 (USP12) is a specialized cysteine protease with a predicted molecular weight of 43 kDa. USP12 is a member of the peptidase C19 family and USP12/USP46 subfamily, and the human protein shares 98% amino acid sequence identity with its mouse ortholog. Like USP1 and USP46, USP12 lacks appreciable activity in the absence of its co-activator UAF1. Addition of UAF1 increases the in vitro activity of USP12 20-fold or more. WDR20 is often found in complex with both USP12 and UAF1, and the ternary complex is reported to be more active than the USP12/UAF1 complex. Biologically, USP12 has been implicated as a positive regulator of Androgen Receptor (AR) activity via a mechanism that involves down-regulation of Akt activity. As such, it has been proposed that USP12 inhibition could offer a therapeutic target for some forms of prostate cancer. Recombinant USP12 contains a C-terminal 6-His tag. Recombinant UAF1 also contains a C-terminal 6-His tag. The assembled complex contains 25 µg USP12 and 45 µg UAF1, resulting in a 1:1 stoichiometry.
References:
Burska, U.L. et al. (2013) J Biol Chem. 288: 32641
Cohn, M.A. et al. (2009) J Biol Chem. 284: 5343
Gangula, N.R. & Maddika, S. (2013) J Biol Chem. 288: 34545
Kee, Y. et al. (2010) J Biol Chem. 285: 11252
McClurg, U.L. et al. (2014) Oncotarget 5: 7081
Ohashi, M. et al. (2015) PLoS Pathog. DOI: 10.1371/journal.ppat.1004822
Long Name:
Ubiquitin Specific Protease 12/USP1-associated Factor 1 Complex
Alternate Names:
USP12/UAF1 Complex
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