Casein is considered to be a generic substrate for a broad spectrum of proteases. As native casein this fluoresceinated casein is hydrolyzed by many proteases, and widely used for fluorimetric measurement of protease activity. In the intact substrate, casein is heavily labeled with 5-FITC, resulting in significant fluorescence quenching. Protease-catalyzed hydrolysis relieves its quenching effect, yielding brightly green fluorescent dye-labeled short peptides. The increase in fluorescence intensity is directly proportional to protease activity. We do not recommend that this conjugate be used for fluorescence polarization assay. For fluorescence polarization we can custom-make the lightly labeled fluorescein casein conjugate.
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