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.

目的：我们旨在评估胺反应性荧光试剂在估计酶促蛋白水解中的潜在应用。

背景：蛋白水解酶在调节生物体的许多生理过程中发挥着重要作用。

目的：通过使用蛋白质测定技术试剂评估蛋白质降解。

方法：我们用 Chromeo P503（Py 1 pyrylium 化合物）和 CBQCA（3-（4-羧基苯甲酰基）喹啉-2-甲醛）作为胺反应试剂和 NanoOrange非胺活性染料。

结果：当用 Chromeo P503（Py 1 pyrylium 化合物）和 CBQCA（3-（4-羧基苯甲酰基）喹啉-2-甲醛进行测定时，所有用胰蛋白酶制备的 BSA 样品都显示出显着高于对照（反映蛋白水解）的荧光强度 (FI) ) 作为胺反应试剂。然而，用 NanoOrange 作为非胺反应试剂测定的相同样品在含有胰蛋白酶的样品和对照样品之间没有显示任何显着差异。

结论：这些结果证实了利用胺反应性荧光试剂对蛋白水解进行一般估计的高灵敏度蛋白质测定的可靠性。