Boc-Val-Pro-Arg-MCA hydrochloride (Boc-Val-Pro-Arg-MCA HCl) is a sensitive fluorescent substrate for the determination of trypsin-like serine proteases activity.

Guidelines (Following is our recommended protocol. This protocol only provides a guideline and should be modified according to your specific needs).
Add 100 μL of diluted Trypsin enzyme to 800 μL of 50 mM Tris-HCl buffer (pH 8.0). Immediately initiate the reaction by adding 100 μL of 50 μM substrate Boc-Val-Pro-Arg-AMC hydrochloride. Incubate at 55°C for 10 minutes. Stop the reaction by adding 1.5 mL of stop solution (methanol: n-butanol: distilled water = 35:30:35, v/v/v). Measure fluorescence intensity using a fluorescence spectrophotometer (excitation 380 nm, emission 450 nm).

Boc-Val-Pro-Arg-MCA hydrochloride participates in the trypsin activity assay
1. Add 100 μL of diluted Trypsin enzyme to a test tube containing 800 μL of 50mM Tris-HCl buffer (pH 8.0).
2. Add 100 μL of 50 μM substrate Boc-Val-Pro-Arg-MCA to start the reaction immediately.
3. Incubate at 55°C for 10 minutes. After incubation, quickly add 1.5 mL of stop agent (methanol: n-butanol: distilled water = 35:30:35) to terminate the reaction.
4. Use a fluorescence spectrometer to measure the fluorescence intensity at an excitation wavelength of 380 nm and an emission wavelength of 450 nm.