[TR-FRET/Kits] XIAP E3 Ligase TR-FRET Kit [SBB-KF0057]_South Bay Bio - 코아사이언스

XIAP E3 Ligase TR-FRET Kit

Cat.# SBB-KF0057 / Size: 400 wells(20μl / well)

 

The kit uses ubiquitin labeled with either Europium-Cryptate or Cyanine5 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 XIAP leads to an increase in fluorescence emission at 665 nm (Emacceptor) and decrease at emission wavelength 620 nm (Emdonor).

Description

South Bay Bio’s XIAP E3 Ligase TR-FRET Kit provides a fast and sensitive method monitoring ubiquitin conjugation onto XIAP 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 XIAP 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 Cyanine5 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 XIAP leads to an increase in fluorescence emission at 665 nm (Emacceptor) and decrease at emission wavelength 620 nm (Emdonor).

Specifications

 

Readout:	Endpoint / Kinetic
Label or Dye:	Europium Cryptate & Cy5
Detection Method:	TR-FRET
Substrate Properties:	Protein-Based Substrate. Typical experimental concentration is 1x.
Excitation/Emission (nm):	304/620 & 304/665
Storage	Store at −80°C after product arrival. Avoid multiple freeze / thaws. It is recommended to make multiple aliquots after the first thaw.

 

Citations & References

1) Magennis, S. W., Parsons, S., Pikramenou, Z., Corval, A., & Woollins, J. D. (1999). Imidodiphosphinate ligands as antenna units in luminescent lanthanide complexes.Chemical communications, (1), 61-62.

2) Zheng, N., & Shabek, N. (2017). Ubiquitin Ligases: Structure, Function, and Regulation.Annual Review of Biochemistry, (0).

 

코아사이언스 coresciences South bay bio 한국 대리점 Ubiquitin proteasome system Real-time TR-FRET conjugation assays cryptases chemistry UB conjugation UB deconjugation C-terminal derivatives UBL derivatives inhibitors cryptate donors cryptate acceptors cryptate kits Di-ubiquitin tetra-ubiquitin immunoproteasomes 20S proteasomes substrates E1s E2s E3s ubiquitinated substrates DUBS