ProteaseSpots™
JPT Peptide Technologies offers a new and efficient way to study protease activities and substrate specificities. ProteaseSpots™ are custom synthesized protease substrates that are immobilized on cellulose discs. The discs are delivered ready-to use in microtiter plates and are incubated with your protease. Subsequent fluorescent detection can be performed after several time points.
Benefits of the ProteaseSpots™ System
- Screen large numbers of substrates economically
- You specify the individual substrate sequences
- Obtain semi-kinetic data
- Easy handling in 96-well plates
Applications for ProteaseSpots™
- Detection of protein cleavage sites utilizing peptide scans
- Identification of novel substrates with de novo libraries
- Measurement of protease activities with known substrates
- Characterization of cleavage sites through substitutional analyses
Figure: The protease assay is based on peptides bound to small cellulose discs in 96-well microtiter plates.
These peptides bear a fluorescent moiety at the N-terminus. After incubation with the active protease, aliquots are transferred to an additional plate for detection of the N-terminally labeled peptide fragment (excitation: 325 nm, emission: 420 nm).
Figure: Fluorescence intensities of a caspase-3 substrate VDQMDGW single site L-amino acid substitutional analysis.
This method identifies new or improved substrates.
Each residue in the sequence (rows) was replaced by all 20 amino acids (columns).
Figure: Fluorescence intensity of trypsin cleaved peptides derived from hen egg lysozyme.
The entire sequence was synthesized using 7-mer overlapping peptides shifted by three amino acids.
Related products and services
Protease Substrate Sets
Internally quenched (Dabcyl/EDANS) pre-made protease substrate collections ready-to-screen in microtiter plates.
Protease Substrate Identification
Profiling service for protease using our high-content PepStar™ peptide microarrays.
Selected References for ProteaseSpots™:
Identification of Candidate Substrates for Ectodomain Shedding by the Metalloprotease-Disintegrin ADAM8
Naus et al., Biol Chem. 2006 (2006)
Screening a Combinatorial Peptide Library to Develop a Human Glandular Kallikrein-2 Activated Prodrug as Targeted Therapy for Prostate Cancer
Janssen et al, Mol. Cancer Ther. (2004)
Processing of the Human Transferrin Receptor at Distinct Positions within the Stalk Region by Neutrophil Elastase and Cathepsin
Kaup et al., Biol. Chem. (2002)
Escherichia Coli DegP Protease Cleaves Between Paired Hydrophobic Residues in a Natural Substrate: the PapA Pilin
Jones et al, J. Bacteriol. (2002)
More references under JPT Publications/Literature
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