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.

The above information is based on published literature. Experimental procedures should be appropriately modified to meet specific research demands.