Recombinant Human His6-USP46/UAF1 Complex Protein, CF

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Summary
Reactivity HuSpecies Glossary
Applications Bioactivity
Format
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Recombinant Human His6-USP46/UAF1 Complex Protein, CF Summary

Details of Functionality

Recombinant Human USP46/UAF1 complex is a Ubiquitin-specific deconjugating enzyme heterodimer.  Reaction conditions will need to be optimized for each specific application.  We recommend an initial USP46/UAF1 Complex concentration of 20-200 nM when using Ubiquitin-AMC or Ubiquitin-Rhodamine (U-550, U-555) as a substrate.  In vitro, USP46/UAF1 complex does not efficiently hydrolyze poly-Ubiquitin chains in the absence of WDR20.

Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human USP46/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
43 kDa (USP46), 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 7.8, 100 mM NaCl, 10% (v/v) Glycerol, 1 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-USP46/UAF1 Complex Protein, CF

  • USP46/UAF1 Complex

Background

Ubiquitin carboxyl-terminal hydrolase 46 (USP46) is a specialized cysteine protease with a predicted molecular weight of 42 kDa. USP46 is a member of the peptidase C19 family and USP12/USP46 subfamily, and the human protein shares 100% amino acid sequence identity with its mouse ortholog. Like USP1 and USP12, USP46 lacks appreciable activity in the absence of its co-activator UAF1. Addition of UAF1 increases the in vitro activity of USP46 10-fold or more.  Biologically, USP46 has been implicated in the regulation of Alpha-Amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid receptors (AMPARs). USP46 is able to deubiquitinate AMPARs both in vivo and in vitro, and by regulating the status of AMPAR ubiquitination and turnover, this deubiquitinase may play an important role in synaptic plasticity and brain function. Recombinant USP46 contains a C-terminal 6-His tag.  Recombinant UAF1 also contains a C-terminal 6-His tag.  The assembled complex contains 25 µg USP46 and 45 µg UAF1, resulting in a 1:1 stoichiometry.

  1. Cohn, M.A. et al. (2009) J Biol Chem. 284: 5343
  2. Huo, Y. et al. (2015) J Neurochem. doi: 10.1111/jnc.13194.
  3. Ohashi, M. et al. (2015) PLoS Pathog.  DOI: 10.1371/journal.ppat.1004822

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