• 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)

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:

1. Burska, U.L. et al. (2013) J Biol Chem. 288: 32641

2. Cohn, M.A. et al. (2009) J Biol Chem. 284: 5343

3. Gangula, N.R. & Maddika, S. (2013) J Biol Chem. 288: 34545

4. Kee, Y. et al. (2010) J Biol Chem. 285: 11252

5. McClurg, U.L. et al. (2014) Oncotarget 5: 7081

6. 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