Ubiquitin Conjugation Initiation Kit
- 貨號 Cat. No.
- K-995
- 規格 Size
- 1 kit
- 品牌 Brand
- Boston Biochem
Description
Ubiquitin Conjugation Initiation Kit
human recombinant
Product Information
1 kit
Use & Storage
Store at -80°C. Avoid multiple freeze/thaw cycles.
Catalog #: K-995
Ubiquitin Conjugation Initiation Kit
Product Source:
human recombinant
This kit is designed for the purpose of charging ubiquitin by E1, the first reaction in the conjugation cascade. This E1-Ubiquitin thioester can be subsequently used for the initiation of any E1 mediated reaction including E2 ubiquitin thioester formation and/or the ubiquitination conjugation reaction of target substrate proteins with the addition of E2 / E3 enzymes (not supplied). The reagents supplied allow for the thioester formation and detection of E1-S-Ub and/ or E2-S-Ub with the addition of E2 enzyme (not supplied).
NOTE: Kit contains reagents sufficient for 10 x 20 μl reactions.
Product Information
Quantity:
1 kit
Structure (deprecated):
| Concentration | Quantity | |
| 20X E1 Enzyme Solution | X mg/ml (X μM) | 15 μl |
| 10X Ubiquitin Solution | X mg/ml | 25 μl |
| 10X Mg-ATP Solution | X mM | 25 μl |
| 10X Reaction Buffer | 0.5 M Hepes pH 8.0 | 25 μl |
| 5X Non-reducing loading buffer | 80 μl |
Background:
The covalent bond between ubiquitin and substrate proteins requires the activation of the C-terminal carboxyl group of Ub to be activated. In this reaction, ubiquitin is linked to the Ubiquitin Activating Enzyme (E1) through a thioester bond between its C-terminal glycine and the active site cysteine
residue in the enzyme (E1-S-Ub). E1 hydrolyzes ATP to AMP and PPi with the intermediate formation of an E1-bound ubiquitin adenylate. The activated Ub is then transferred from E1 to any of a family of Ubiquitin Carrier Protein Enzymes (E2s) in a similar thioester linkage (E2-S-Ub). Ubiquitin can then be transferred directly to the ε-Lys of a substrate from E2 enzymes. Other conjugation reactions require the involvement of a third enzyme, Ubiquitin Ligase (E3). E3 binds to substrate proteins and also to an E2 to form an E2-E3-substrate complex. It is the recognition and formation of said complex that has the highest level of substrate specificity for the conjugation
cascade.
Use & Storage
Storage:
Store at -80°C. Avoid multiple freeze/thaw cycles.