Aims: We aim to evaluate the potential application of amine reactive fluorogenic reagents
for estimating enzymatic proteolysis.
Background: Proteolytic enzymes play important roles in regulating many physiological processes
in living organisms.
Objectives: Assessment of protein degradation by using reagents for protein assay techniques.
Methods: We have assayed samples at the start and after 30-60 minutes incubation with trypsin by
Chromeo P503 (Py 1 pyrylium compound) and CBQCA (3-(4-carboxybenzoyl) quinoline-2-carboxaldehyde)
as amine reactive reagents and NanoOrange as non-amine reactive dye.
Results: All BSA prepared samples with trypsin have shown significantly higher fluorescence intensity
(FI) versus controls (which reflects proteolysis) when assayed by Chromeo P503 (Py 1
pyrylium compound) and CBQCA (3-(4-carboxybenzoyl) quinoline-2-carboxaldehyde) as amine reactive
reagents. However, same samples assayed with NanoOrange as non-amine reactive reagent
did not show any significant variation between samples containing trypsin and controls.
Conclusion: These results are confirming reliability of highly sensitive protein assays utilizing
amine reactive fluorogenic reagents for general estimation of proteolysis.