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Proteases are widely present in animal viscera, plant stems, leaves, fruits, and microorganisms. Acidic protease (ACPT) hydrolyzes casein to produce tyrosine under acidic conditions; tyrosine reacts with Folin-Ciocalteu reagent to form a blue compound; this blue substance has a characteristic absorption peak at 680nm, allowing for the determination of acidic protease activity. Since the substrate casein itself contains various amino acids, a control with casein must be set during the detection process to subtract interfering background values and exclude false positives.
Additional Materials and Equipments Required
Microplate reader, 96-well plate, water bath, magnetic stirrer, pipette, distilled water.
