Material Data Sheet

Ubiquitin Conjugation Initiation Kit

Price: $119.00
Catalog #: K-995

Ubiquitin Conjugation Initiation Kit

1 Kit
Data Sheet: 

This kit is designed for the purpose of charging (activating) Ubiquitin with an E1 Ubiquitin Activating Enzyme, the first reaction in the Ubiquitin conjugation cascade.  This E1-Ubiquitin thioester can be subsequently used for the initiation of E1-mediated in vitro reactions including E2-Ubiquitin thioester formation and the ubiquitination of target substrate proteins with the addition of appropriate E2 and E3 Ubiquitin Ligase enzymes (not supplied).

NOTE: Kit contains reagents sufficient for 12 x 20 μl reactions.

Product Information

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.

Store at -80°C. Avoid multiple freeze/thaw cycles.