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.

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

DMSO: 50 mg/mL (74.95 mM), Sonication is recommended.