Description
ChoosE2-Opti™ Intrachain TR-FRET Assay Kit | 78561 | Gentaur US, UK & Europe Disrtribition
Category: Ubiquitination
Application: Identify E2 enzymes that are partners of an E3 ligase of interest Assess the activity of new E2-E3 protein pairs Screen new E3 ligases and variants in HTS applications Screen molecules that inhibit/activate E2 enzyme activity in HTS applications, Determine compound IC50 Analyze E3 ligase activity in real-time
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 largely determined by the E2-E3 complex where the E3 enzyme directs the last step of the Ub-conjugating cascade by binding to both an E2~Ub conjugate and a substrate protein.
Description: ChoosE2-Opti™ Intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit, designed to identify E2 enzyme(s) that are functional partners for a purified E3 ubiquitin ligase of interest and lead to its self-polyubiquitination. The assay kit contains five E2 enzymes to be tested with the E3 ligase of interest, and two E3 ligases (SMURF1 and MDM2) to use as internal controls. It also contains an E1 enzyme, ATP, an optimized TRF Ubiquitin Mix, and a universal buffer. It utilizes a Europium-labeled Ubiquitin Donor as well as Cy5-labeled Ubiquitin Acceptor to complete the TR-FRET pairing. Since both the TR-FRET donor and acceptor are incorporated into polyubiquitin chains, this FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetics analyses.
Product Type: Assay Kit
Shippement Condition: -80°C
