Ubiquitin-Propargylamide

Catalog number: SBB-PS0034, 50 μg

Ubiquitin- Propargylamideis a potent and specific inhibitor of deubiquitinating enzymes (DUBs) based on a C-terminal electrophilic Propargylamidegroup. Ub- Propargylamidecan be used for activity profiling experiments and DUB inhibitor specificity determination. It targets four of the five major DUB families: UCH, USP, OUT, and MJD, while leaving JAMM metalloproteases uninhibited. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human Ubiquitin-Propargylamide concentration of 1-5 μM.

In stock
SKU
0034
$135.00

Description

Ubiquitin is a small (8.5 kDa) regulatory protein that has been found in almost all tissues of eukaryotic organisms. The addition of ubiquitin to a substrate protein is called ubiquitination or ubiquitylation. Ubiquitination can affect proteins in many ways; it can signal for their degradation via the proteasome, alter their cellular location, affect their activity, and promote or prevent protein interactions. Removal of ubiquitin from a substrate protein occurs via deconjugating enzymes, of which there are nearly 100 known enzymes with various linkage specificities. This product consists of a full-length human, mature ubiquitin polypeptide (amino acids 1-76), expressed in E.coli with a c-terminal warhead (Propargylamide). Ubiquitin-Propargylamide is a potent, irreversible, and specific inhibitor of deubiquitinating enzymes (DUBs) based on a C-terminal electrophilic Propargylamide group. Ub-Propargylamide can be used for activity profiling experiments and determining DUB inhibitor specificity. It targets three of the four major DUB families: UCH (Ubiquitin c-terminal hydrolases), USP (Ubiquitin Specific Proteases), OTU (Ovarian Tumor Proteases), and MJD (Machado-Josephin domain proteases) while JAMM metalloproteases are not inhibited. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human Ubiquitin Propargylamide concentration of 1-5 μM.

Specifications

Product Overview
Quantity: 50 μg
Molecular Weight: 8.6 kDa
Purity: >97% by LCMS
Label or Dye: Propargylamide
Substrate Properties: Protein-Based Substrate. Typical experimental concentration 1-5 μM.
Storage Buffer: 50mM MES pH 6.0, 100mM NaCl
Storage Store at −80°C after product arrival. Avoid multiple freeze / thaws. It is recommended to make multiple aliquots after the first thaw.

Figures & Data

UCHL3 + Ub-AMC + Ubiquitin-Propargylamide

Figure 1. Kinetic Activity. UCHL3 (SBB-DE0023) activity with Ubiquitin-AMC (SBB-PS0043) measured in the presence of 15.0 nM and 0.1 nM Ubiquitin-Propargylamide.

Citations & References

1) Borodovsky et al. 2001, 20, 5187.

2) Xu et al. RSC Adv 2016, 6, 
47926. 


3) Tong, X. et al. (2012) J. Biol. Chem. 287:25280. Galardy et al. Methods in Enzymology 2005, 399, 120.