JPT Peptide Technologies offers an 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

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
Peptide Microarray Assay Service
Profiling service using high content PepStar peptide microarrays

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