JPT Peptide Technologies offers a new and efficient way to study protease activities and substrate specificities. ProteaseSpots™ are custom synthesized protease substrate peptides that are immobilized on cellulose discs. The discs are delivered ready-to use in microtiter plates. During incubation with protease samples fluorescent detection can be performed after several time points.
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).
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
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
Selected References for ProteaseSpots™
"Identification of Candidate Substrates for Ectodomain Shedding by the Metalloprotease-Disintegrin ADAM8"
Naus et al., Biol Chem. 2006 (2006) - PMID: 16542157
"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) - PMID:15542783
"Processing of the Human Transferrin Receptor at Distinct Positions within the Stalk Region by Neutrophil Elastase and Cathepsin"
Kaup et al., Biol. Chem. (2002) - PMID: 12222675
"Escherichia Coli DegP Protease Cleaves Between Paired Hydrophobic Residues in a Natural Substrate: the PapA Pilin"
Jones et al, J. Bacteriol. (2002) - PMID: 12270835
Read More References with ProteaseSpots™
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.
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