Description
ChoosE3-Freedom™ Intrachain TR-FRET Assay Kit | 78560 | Gentaur US, UK & Europe Disrtribition
Category: Ubiquitination
Application: Measure the auto-polyubiquitination of any E3 ligase of interest Identify novel E3 ligases or variants of known ligases Screen inhibitors or activators of an E3 ligase of interest in HTS applications Determine compound IC50 Perform E3 ligase real-time analyses
Background: Covalent conjugation to ubiquitin (Ub) is one of the major post-translational modifications that regulates protein stability, function, and localization. Ubiquitination is the concerted action of three enzymes: a Ub-activating enzyme (E1), a Ub-conjugating enzyme (E2), and a Ub ligase (E3). The specificity and efficiency of ubiquitination are determined by the E3 enzyme, which directs the last step of the Ub-conjugating cascade by binding to both an E2~Ub conjugate and a substrate protein. This step ensures the transfer of Ub from E2~Ub to the substrate, leading to its mono- or poly-ubiquitination.
Description: The ChoosE3-Freedom Intrachain TR-FRET Assay Kit contains E1 and E2 enzymes, ATP, an optimized TRF Ubiquitin Mix, and a universal buffer. Purified E3 ligase MDM2 is also provided as an internal quality control. The assay was designed with a Europium-labeled Ubiquitin donor and a Cy5-labeled Ubiquitin acceptor to complete the TR-FRET pairing. Since both the TR-FRET donor and acceptor are incorporated into poly-ubiquitin chains formed on the E3 ligase, the assay measures only poly-ubiquitination and not mono-ubiquitination. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetics.
Product Type: Assay Kit
Shippement Condition: -80°C
