Recombinant Human His6-USP46/UAF1 Complex Protein, CF Summary
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.
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
E-586
| Formulation | X mg/ml (X μM) in 50 mM HEPES pH 7.8, 100 mM NaCl, 10% (v/v) Glycerol, 1 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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Reconstitution Calculator
Background: USP46/UAF1 Complex
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.
- Cohn, M.A. et al. (2009) J Biol Chem. 284: 5343
- Huo, Y. et al. (2015) J Neurochem. doi: 10.1111/jnc.13194.
- Ohashi, M. et al. (2015) PLoS Pathog. DOI: 10.1371/journal.ppat.1004822
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The experiment was done with U-555
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