Recombinant Human His6-USP30 Protein, CF Summary
Reaction conditions will need to be optimized for each specific application. We recommend an initial USP30 concentration of 20-100 nM in reactions utilizing Ubiquitin-AMC or Ubiquitin-Rhodamine substrates (U-550 or U-555), or 200-500 nM when digesting recombinant Polyubiquitin chain substrates.
Contains a C-terminal 6-His tag, Thr57 - Glu517
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.
|Formulation||X mg/ml (X μM) in 50 mM HEPES pH 7.2, 0.4 M NaCl, 10% Glycerol (v/v), 2 mM DTT|
|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.
Ubiquitin carboxyl-terminal hydrolase 30 (USP30) is a member of the C19 family of peptidases and is tethered to the outer mitochondrial surface via a trans-membrane domain. USP30 activity is antagonistic to the pro-mitophagy signal generated by the activities of E3 Ubiquitin Ligase Parkin (PARK2) and kinase PINK1, which are stimulated by mitochondrial damage. In vivo, USP30 cleaves Ubiquitin from mitochondrial surface proteins such as MIRO1, TOMM20, MFN1, and MFN2. The enzyme displays a strong preference for K6- and K11-linked Polyubiquitin chains in vitro, but has substantially lower activity on Polyubiquitin chains that have been phosphorylated with PINK1. This recombinant protein contains amino acids 57-517 of the full-length protein, and a C-terminal 6-His tag.
- Bingol B. et al. (2014) Nature 510: 370
- Cunningham C.N. et al. (2015) Nat Cell Biol. 17: 160
- Thobois S. (2015) Mov. Disord. 30: 340
- Wang, Y. et al. (2015) Autophagy 11: 595
- Wauer T., et al. (2015) EMBO J. 34: 307
- Yue, W. et al. (2014) Cell Res. 24: 482
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