CHOP Isopeptidase Assay

CHOP: a revolutionary technology for characterizing
UB /UBL isopeptidase
activity

CHOP-Reporter technology offers a rapid and robust in vitro assay for Ubiquitin and Ubiquitin-like Isopeptidase activity. Previous Ub/Ubl Isopeptidase assays relied on the fusion of small chemical adducts to the C-terminus of the Ub/Ubl. In many cases this molecule is not recognized by the Isopeptidase of interest. The CHOP assay platforms utilize protein fusions which are more physiologically relevant and can be recognized by Isopeptidases that do not recognize other substrates.

Assay Principle

The concept behind the Ub/Ubl-CHOP Reporter Deconjugation Assay is the fusion of Ub/Ubl with reporter enzyme. The nature of this fusion renders the reporter enzyme catalytically inactive. Upon cleavage of the Ub/Ubl-fusion by an isopeptidase, the activated, free reporter enzyme acts upon its substrate and generates a fluorescent signal. Thus, in the coupled assay, the signal generated by cleavage of the reporter enzyme’s substrate is a quantitative measure of isopeptidase activity.
CHOP and CHOP2 platforms utilize different reporter enzymes making multiplexing possible.

High sensitivity of CHOP Assays

Range of Kits

ISG15-CHOP-Reporter Kit SUMO3-CHOP-Reporter Kit NEDD8-CHOP2-Reporter Kit
NEDD8-CHOP-Reporter Kit Ub-CHOP-Reporter Kit SUMO3-CHOP2-Reporter Kit
Smt3-CHOP-Reporter Kit ISG15-CHOP2-Reporter Kit Ub-CHOP2-Reporter Kit

 

 

 

 

SensoLyte® Deubiquitination Assay Kits

The SensoLyte® 520 Deubiquitination Assay Kit provides a convenient assay for inhibitors/activators screening in deubiquitinylation or for continuous assay of enzyme activity using a fluorogenic peptide. Upon protease cleavage, this substrate generates the bright green fluorophore that can be detected at excitation/emission=490/520 nm. The long wavelength spectra of the green fluorophore released after cleavage of the substrate provides less interference from other reaction components.
 
The SensoLyte® 440 Deubiquitination Assay Kit provides a convenient assay for inhibitors/activators screening in deubiquitinylation or for continuous assay of enzyme activity. A natural substrate, ubiquitin, coupled with the AMC (7-amino-4-methylcoumarin) fluorophore is used in the kit. Upon the cleavage by a deubiquitin protease, this substrate generates the blue fluorophore that can be detected at excitation/emission=354/442 nm. The substrate supplied in the kit is exquisitely sensitive for ubiquitin hydrolases (UCHs).
 
 
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