ChoosE3-Freedom™ Intrachain TR-FRET Assay Kit

TheChoosE3-Freedom Intrachain TR-FRET Assay Kitis a sensitive high-throughput screening (HTS) TR-FRETassaykit, designed tomeasuretheauto-polyubiquitination ofanypurifiedE3 ligaseof interestin a homogeneous 384 reaction format.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.Figure 1: ChoosE3-Freedom Intrachain TR-FRET Assay Kit schematic.Need us to run inhibitor screens or profile your compounds against ChoosE3-Freedom™? Check out ourUbiquitination Screening ServicesorDNA Replication & Repair Screening Services.