% Signal to Background of Continuous Real-Time TR-FRET Parkin titration (autoubiquitination): Serial dilutions of Parkin W403A from 50nM to 3.125nM and 300nM wt Parkin were mixed with UBA1, UBE2D3 and trf-Ub (pS65) mix. Reactions were initiated wit
% Signal to Background of Continuous Real-Time TR-FRET Parkin titration (autoubiquitination): Serial dilutions of Parkin W403A from 50nM to 3.125nM and 300nM wt Parkin were mixed with UBA1, UBE2D3 and trf-Ub (pS65) mix. Reactions were initiated wit
Parkin E3 Ligase TR-FRET Kit
SBB-KF0036
Product group Chemicals
Overview
- SupplierSouth Bay Bio
- Product NameParkin E3 Ligase TR-FRET Kit
- Delivery Days Customer10
- CertificationResearch Use Only
- Hazard InformationNon-hazardous
- Scientific DescriptionSouth Bay Bios Parkin TR-FRET Ubiquitin Kit provides a fast and sensitive method monitoring ubiquitin conjugation onto both wild-type Parkin and a more active mutant W403A in solution, resulting from an enzymatic ubiquitin cascade without the need of running and staining an SDS gel. The kit enables continuous TR-FRET detection of ubiquitin chain formation onto Parkin in a real-time detection setup, or in an end-point configuration if desired. TR-FRET uses the extended fluorescence emission decay lifetimes typical of rare-earth lanthanides to impart a short time-delay between FRET donor excitation and emission. This delay provides a means to separate true signal from short-lived background fluorescence, and reduce interference from compound fluorescence and other assay artifacts. - South Bay Bios Parkin TR-FRET Ubiquitin Kit provides a fast and sensitive method monitoring ubiquitin conjugation onto both wild-type Parkin and a more active mutant W403A in solution, resulting from an enzymatic ubiquitin cascade without the need of running and staining an SDS gel. The kit enables continuous TR-FRET detection of ubiquitin chain formation onto Parkin in a real-time detection setup, or in an end-point configuration if desired. TR-FRET uses the extended fluorescence emission decay lifetimes typical of rare-earth lanthanides to impart a short time-delay between FRET donor excitation and emission. This delay provides a means to separate true signal from short-lived background fluorescence, and reduce interference from compound fluorescence and other assay artifacts. The kit uses ubiquitin labeled with either Europium-Cryptate or Cy5 as FRET pair donor and acceptor fluorophores respectively, completely eliminating the need for antibody based detection setups. Enzymatic incorporation of the labeled ubiquitins into chains conjugated onto Parkin leads to an increase in fluorescence emission at 665nm (Em-Acceptor) and decrease at emission wavelength 620nm (Em-Donor).
- Storage Instruction-20°C,-80°C
- UNSPSC12352200