Ubiquitin-Rhodamine 110

Produktübersicht

• Artikelname: Ubiquitin-Rhodamine 110
• Artikelnummer: UBI-60-0117-050
• Hersteller Artikelnummer: 60-0117-050
• Alternativnummer: UBI-60-0117-050
• Hersteller: Ubiquigent
• Kategorie: Sonstiges
• Spezies Reaktivität: Human

Zusatzinformationen

• Species

  human

• Source

  synthetic

• Quantity

  50 µg

• Storage

  -70°C

• Concentration

  2mg/ml

• Formulation

  DMSO

• Molecular Weight

  8.93kDa

• Stability

  12 months at -70°C; aliquot as required

• Protein Sequence

  Accession number: P62987. C-terminally tagged with Rhodamine 110

• QA; Protein Identification

  Confirmed by mass spectrometry.

• QA; Activity

  Activity Assay: The activity of Ubiquitin-Rhodamine 110 was validated by determining the increase in fluorescence at 535nm (Excitation 485nm) measured as a result of the enzyme catalysed cleavage at the amide bond between the C-terminal Glycine and Rhodamine, generating Ubiquitin and de- quenched Rhodamine 110-Glycine. UCHL3 (deubiquity-lase) was incubated with Ubiquitin-Rhodamine 110 and the fluorescence was measured at four time points (0min, 30min, 60min and 90min).

 

Background:

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).

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.

Produkteigenschaften

• Konzentration: 2mg/ml
• Molekulargewicht: 8.93kDa
• NCBI: <a href=https://www.ncbi.nlm.nih.gov/gene/P62987 target=_blank rel=nofollow>P62987
• Quelle: synthetic
• Sequenz: Accession number: P62987. C-terminally tagged with Rhodamine 110