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).

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

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.

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