Boc-Val-Pro-Arg-MCA hydrochloride is a sensitive fluorogenic substrate for measuring trypsin-like serine proteases activity. In Vitro Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). Trypsin activity assay1. Add diluted Trypsin enzyme (100 μL) to 800 μL of 50 mM Tris–HCl buffer (pH 8.0). 2. The reaction is immediately initiated by the addition of 100 μL of 50 μM substrate Boc-Val-Pro-Arg-MCA. 3. Incubate at 55 °C for 10 min. 4. Add 1.5 mL of the stopping agent (methyl alcohol:n-butyl alcohol:distilled water = 35:30:35, v/v/v) to stop the reaction. 5. Measure the fluorescence intensity by a spectrofluorometer (excitation 380 nm, emission 450 nm). MCE has not independently confirmed the accuracy of these methods. They are for reference only. Form:Solid