Recombinant Human His6-USP12/UAF1 Complex Protein, CF

R&D Systems | Catalog # E-580

R&D Systems
Discontinued Product
E-580 has been discontinued. View all USP12/UAF1 Complex products.

Key Product Details

Source

Sf 21 (baculovirus)

Applications

Bioactivity
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Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human USP12/UAF1 Complex protein
Contains a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Predicted Molecular Mass

44 kDa (USP12), 77 kDa (UAF1)

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.

Reviewed Applications

Read 1 review rated 5 using E-580 in the following applications:

Formulation, Preparation, and Storage

E-580
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.
  • 12 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

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

Product Documents for Recombinant Human His6-USP12/UAF1 Complex Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human His6-USP12/UAF1 Complex Protein, CF

For research use only

Customer Reviews for Recombinant Human His6-USP12/UAF1 Complex Protein, CF (1)

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    Recombinant Human His6-USP12/UAF1 Complex Protein, CF E-580

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