The Sortase A Assay Kit is a fluorogenic assay kit designed to measure Sortase A proteolytic activity for screening and profiling applications.
Figure 1: Illustration of the assay principle.
The fluorescence emitted by donor DNP is quenched due to the proximity of the Abz acceptor in the intact peptide. The protease cle*es the peptide between T (threonine) and G (glycine) and forms an intermediate complex between the cysteine residue within the active site and the C-terminus of the peptide substrate. The cle*ed substrate is then covalently bounded to the triglycine nucleophile. The glycine-DNP fragment is released, freeing the emission of fluorescence by DNP.
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