Parkin Auto-Ubiquitination Kit
- 貨號 Cat. No.
- K-105
- 規格 Size
- 1 kit
- 品牌 Brand
- Boston Biochem
Description
Parkin Auto-Ubiquitination Kit
human recombinant
Product Information
1 kit
Use & Storage
Store at -80°C. Avoid multiple freeze/thaw cycles.
Catalog #: K-105
Parkin Auto-Ubiquitination Kit
Product Source:
human recombinant
This kit is designed as a control for to the conjugation of the ubiquitin to protein substrates in vitro, which requires the activities of the ubiquitin E1 (E-305), E2 UbcH7 (E2-640) and E3 Parkin (E3-150) activating enzyme. The E1 enzyme charges the ubiquitin by forming an ATP-dependent high energy thiolester bond. The activated ubiquitin is subsequently transferred to UbcH7 then to Parkin. The Parkin-S-Ub complex has the ability to both auto-ubiquitinate and transfer the ubiquitin to various substrates such as CDCrel-1 and rel-2a, cyclin E, synphilin-1, the O-glycosylated form of α-synuclein (αSp22), PAel-R, FBP1, α/β tubulin, RanBP2, Hsp70, synaptotagmin XI. Other E2s that have shown activity with Parkin are UbcH8 (E2-644) and UbcH13/Uve1a (E2-664). Alternatively labeled ubiquitins may be substituted for biotin-ubiquitin for visualization such as fluorescein-ubiquitin (U-590) and rhodamine-ubiquitin (U-600).
NOTE: Kit contains reagents sufficient for 10 x 20 μl reactions.
Concentration of components vary with Lot #.
Product Information
Quantity:
1 kit
Supplied:
| Concentration | Volume | |
| 10X E1 Enzyme | X mg/ml (X μM) | 20 μl |
| 10X UbcH7 | X mg/ml (X μM) | 20 μl |
| 10X His6-Parkin | X mg/ml (X μM) | 20 μl |
| 10X Biotin-Ubiquitin | X mM | 20 μl |
| 10X Reaction Buffer | X mM | 20 μl |
Background:
Mutations in the Parkin (PRKN2) gene are considered to be a major cause of autosomal recessive juvenile parkinsonism (AR-PJ). Parkin functions as an E3 ligase having an N-terminal ubiquitin-like motif and a C-terminal RING domain composed of two RING finger motifs separated by two IBR domains. Parkin can auto-ubiquitination itself and ubiquitinate various substrates (eg. CDCrel-1 and rel-2a, cyclin E, synphilin-1, the O-glycosylated form of α-synuclein (αSp22), PAel-R, FBP1, α/β tubulin, RanBP2, Hsp70, synaptotagmin XI) in an E2-dependent manner, targeting them for degradation. Parkin functions in conjunction E2 enzymes UbcH7 (E2-640), UbcH8 (E2-644) and UbcH13/Uev1 (E2-664). Parkin disease-associated mutations often affect E3 ligase activity through decreased E2 and/or substrate interactions which may thus result in the dysfunction of proteasomal degradation pathways and the neurotoxic accumulation of misfolded proteins.
Use & Storage
Storage:
Store at -80°C. Avoid multiple freeze/thaw cycles.