Europium-Cryptate Ubiquitin, human recombinant

Catalog number: SBB-TR0014, 20 μg

Highly purified recombinant protein site-specifically labeled with a single Europium Cryptate moiety. All lysines are available along with a fully functional C-terminus, making the Europium Cryptate Ub ideal for chain formation using the the E1 – E2 and E3 conjugation cascade. The Cryptate has an excitation maximum of 304nm and an emission maximum of 620nm. This protein was expressed in E.coli.[1][2][3][4]

In stock
SKU
0014
$495.00

Description

Ubiquitin is a highly conserved protein that plays a major role in the ubiquitination pathway, which is conserved from yeast to mammals. Ubiquitination, the conjugation of ubiquitin to other proteins through a covalent bond between its C-terminal glycine and the ε-amino group of lysine residues or the α-amino group of an N- terminal methionine onto proteins is essential for many cellular process primarily linked to protein degradation. This process involves three steps with specific groups of enzymes in an ATP depended manner, which are activation with ubiquitin-activating enzymes (E1s), conjugation with ubiquitin-conjugating enzymes (E2s), and ligation with ubiquitin ligases (E3s). This Europium Cryptate Ubiquitin is ideal for measuring Ub chain conjugation or deconjugation using TR-FRET measurement. Its TR-FRET pair is Cy5-Ubiquitin. Typical working concentrations range from 250 to 500 nM. However, the best working concentration needs to be empirically determined by the type of application by the end user.

For Research Use Only, Not For Use In Humans.

Specifications

Product Overview
Quantity: 20 μg
Molecular Weight: 9.9 kDa
Purity: >98% by LCMS
Readout: Endpoint / Kinetic
Label or Dye: Europium3+ Cryptate
Detection Method: Fluorescence
Substrate Properties: Protein-Based TR-FRET Pair Donor, Typical working concentrations range from 250 to 500 nM. However, the best working concentration needs to be empirically determined by the type of application by the end user.
Excitation/Emission (nm): 304/620
Storage Buffer: 50 mM HEPES pH7.5, 50 mM NaCl
Storage Store at −80°C, avoid multiple freeze-thaws. Make multiple aliquots after the first thaw.

Figures & Data

Mass Spectrometry

Figure 1. LCMS. Analysis of Eu3+ Cryptate-Ubiquitin using LCMS intact mass determination indicates purity greater than 98%, and a molecular weight of 9,855 daltons.

Citations & References

1) Hemmilä, Ilkka and Stuart Webb. "Time-Resolved Fluorometry: An Overview Of The Labels And Core Technologies For Drug Screening Applications". N.p., 2017. Print.

2) Komander, David and Michael Rape. "The Ubiquitin Code". Annual Review of Biochemistry 81.1 (2012): 203-229. Web. 9 Mar. 2017.

3) Visser, A. J. W. G. et al. "Time-Resolved FRET Fluorescence Spectroscopy Of Visible Fluorescent Protein Pairs". European Biophysics Journal 39.2 (2009): 241-253. Web. 13 Mar. 2017.