In addition to fusion proteins, ubiquitin derivatives conjugated with a fluorophore have been reported as substrates for biochemical DUB assays. Ubiquitin-Rhodamine 110 (Ub-Rho110-G) is a fluorogenic rhodamine-based substrate. While the disubstituted rhodamine moiety in UbRho110-G is essentially non-fluorescent, cleavage results in a mono-substituted rhodamine, Rho110-G, which exhibits intense fluorescence when excited at 485 nm (Hassiepen et al., 2007). The rhodamine fluorophore exhibits optical properties more appropriate – than Ubiquitin-AMC – for compound screening and profiling. The risk of artifacts in screens due to autofluorescence of compounds is substantially reduced as the rhodamine 110 fluorophore has excitation and emission wavelengths of 485nm and 535nm respectively (Hassiepen et al., 2007). This product contains Ubiquitin-Rhodamine 110 supplied as a 4x concentrated solution in Ubiquigent’s DUB Assay Buffer. This product is designed to be diluted to 1x (100 nM final concentration) within the assay.

References:

Hassiepen U, Eidhoff U, Meder G, Bulber JF, Hein A, Bodendorf U, et al. (2007) A sensitive fluorescence intensity assay for deubiquitinating proteases using ubiquitin-rhodamine110-glycine as substrate. Anal Biochem 371, 201-207.

Ubiquigent Limited is a specialist developer and supplier of Drug Discovery Services, high quality Research Tools and Chemistry to the life science research community worldwide.

Ubiquigent’s scientific and business interests have a clear focus; namely the ubiquitin, ubiquitin-like, and integrated signalling systems.

The Company has established its scientific and business credentials with both academic researchers undertaking fundamental scientific discovery and pharmaceutical and biotechnology company scientists exploring the potential of ubiquitin cascade-focused drug discovery.

Ubiquigent benefits from high calibre backing including from private US investors, the UK Medical Research Council, and the University of Dundee. The Company’s headquarters and laboratory operations are based in the UK and are located in a state-of-the-art facility adjacent to the MRC Protein Phosphorylation and Ubiquitylation Unit at the University of Dundee (both founded by Professor Sir Philip Cohen). In addition to Ubiquigent’s own facilities and capabilities, such proximity provides ready access to a huge range of additional scientific expertise (1,000+ life-science researchers on site), technological competencies, and assay and analytical platforms.

Ubiquigent staff have a wealth of scientific experience, technological competencies and commercial expertise, gained from within academia, the life science research reagents and services sector, and the pharmaceutical industry, that enables the Company to offer a level of understanding and service to its customers and clients worldwide that few companies may match.