The present invention relates to methods of measuring the activity of a
hydrolytic agent comprising contacting a biomolecule with a hydrolytic
agent in the presence of a fluorescent dye under conditions that allow
digestion of the biomolecule by the hydrolytic agent. The fluorescence of
the dye is monitored over time and a change in fluorescence signifies
digestion of the biomolecule by the hydrolytic agent. The biomolecule is
preferably a protein, peptide or proteome but can also be a carbohydrate,
oligonucleotide or lipid. Further methods relate to determining an end
point for digestion of a biomolecule by a hydrolytic agent, and methods
of monitoring digestion of a biomolecule by a hydrolytic agent. The
monitoring can be performed on the reaction mixture in real time or via
sampling. The invention also relates to kits for carrying out the method.