Recombinant Human His6-USP12/UAF1 Complex Protein, CF

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Summary
Reactivity HuSpecies Glossary
Applications Bioactivity
Format
Carrier-Free

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Recombinant Human His6-USP12/UAF1 Complex Protein, CF Summary

Details of Functionality

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 human USP12/UAF1 Complex protein
Contains a C-terminal 6-His tag
Accession #
Protein/Peptide Type
Recombinant Enzymes
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Applications/Dilutions

Theoretical MW
44 kDa (USP12), 77 kDa (UAF1).
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

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.
Buffer

X mg/ml (X μM) in 50 mM HEPES pH 8.0, 150 mM NaCl, 10% (v/v) Glycerol, 2 mM TCEP

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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human His6-USP12/UAF1 Complex Protein, CF

  • USP12/UAF1 Complex

Background

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.

  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

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