UBCH5b TR-FRET Assay Kit

The Ubch5b TR-FRET Assay Kit is a homogeneous, sensitive TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) assay kit designed to measure UbcH5b (Ubiquitin-conjugating enzyme E2 D2) ubiquitination activity. It utilizes biotin-labeled Ubiquitin and a Terbium-labeled antibody recognizing the His-tagged Ubch5b protein to complete the TR-FRET pairing. The kit contains enough purified UbcH5b, purified UBE1, Biotin-Ubiquitin, anti-His Tb-labeled donor, dye-labeled acceptor, and assay buffer for 400 reactions.Figure 1: UbcH5b TR-FRET Assay Kit schematic.The Terbium-labeled anti-His antibody binds to the His-tagged E2 conjugating protein, while the Dye-labeled streptavidin acceptor binds to Biotin-Ubiquitin. The complex forms when ubiquitin is transferred to the E2 enzyme, and the TR-FRET signal is measured using a fluorescence plate reader capable of measuring Time Resolved-Fluorescence Resonance Energy Transfer.   The TR-FRET signal is proportional to UbcH5b activity.*NOTE: As of January 2024, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.Need us to run inhibitor screens or profile your compounds against UBCH5b? Check out ourUbiquitination Screening Services.