Boc-Val-Pro-Arg-MCA TFA is a sensitive fluorescent substrate that is a thrombin substrate for the determination of trypsin-like serine protease activity.

Boc-Val-Pro-Arg-MCA TFA participates in the activity assay of trypsin
Operation steps
1. Add 100 μL of diluted Trypsin enzyme to a test tube containing 800 μL of 50 mM Tris-HCl buffer (pH 8.0).
2. Add 100 μL of 50 μM substrate Boc-Val-Pro-Arg-MCA TFA to start the reaction immediately. Then incubate at 55°C for 10 minutes.
3. 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.