Boc-Gln-Ala-Arg-AMC hydrochloride is a fluorescent compound used as a substrate for trypsin and for assessing the protein hydrolysis activity of TMPRSS2.

Instructions
1. Solvent selection: Boc-Gln-Ala-Arg-AMC hydrochloride is soluble in dimethyl sulfoxide (DMSO) and water. Working solutions are usually prepared in DMSO at a concentration of 1–10 mM, and the specific concentration is adjusted according to experimental requirements.
2. Substrate preparation: Dissolve Boc-Gln-Ala-Arg-AMC hydrochloride in a suitable solvent (such as DMSO) to prepare a working solution. The concentration of the substrate is usually in the range of 10–100 µM.
3. Enzyme activity detection:
a. Prepare the reaction solution using a buffer with an appropriate pH (usually 7.4), such as phosphate buffered saline (PBS).
b. Add the substrate to the enzyme reaction system, usually at a substrate concentration of 10–50 µM.
c. Incubate the reaction solution and enzyme (such as trypsin or TMPRSS2) at a suitable temperature (usually 37°C).
d. After the enzyme catalyzes the reaction, the substrate is hydrolyzed to generate the fluorescent product AMC (7-amino-4-methylcoumarin).
4. Detection: The hydrolysis of Boc-Gln-Ala-Arg-AMC generates the fluorescent product AMC.
Excitation wavelength: 380 nm
Emission wavelength: 460 nm
The fluorescence intensity can be measured by a fluorescence microplate reader, spectrophotometer or fluorescence microscope to quantitatively analyze the enzyme activity.
Notes:
1) Ensure that the enzyme concentration is optimized to avoid reaction saturation.
2) Maintain appropriate pH and temperature conditions during the experiment to ensure enzyme activity.