4-Methylumbelliferyl heptanoate (4-Methylumbelliferyl enanthate) is a lipase fluorescent substrate that detects cell-mediated cytotoxicity and cell proliferation.

Instructions
1. Solvent selection: 4-Methylumbelliferyl heptanoate is fat-soluble and is usually dissolved in organic solvents such as ethanol, dimethyl sulfoxide (DMSO), etc. The commonly used solution concentration is 1–10 mM, and the specific concentration is adjusted according to the experimental requirements.
2. Substrate addition: Add the 4-Methylumbelliferyl heptanoate solution to the reaction system containing lipase or other target enzymes. Under the catalytic action of lipase, the substrate will be hydrolyzed to generate a fluorescent product.
3. Reaction conditions: The reaction is usually carried out in a buffer in the pH range of 7–8, and the temperature is usually room temperature or 37°C. The reaction time can be adjusted according to the lipase activity and experimental requirements, usually 10–60 minutes.
4. Product detection: The generated fluorescent product can be detected by a fluorescence spectrophotometer with an excitation wavelength of 360 nm and an emission wavelength of 450 nm. Fluorescence signals in cells can also be quantitatively analyzed using a fluorescence microscope or flow cytometer.
5. Cell proliferation and cytotoxicity assays: 4-Methylumbelliferyl heptanoate is also used in cell experiments as a tool for detecting cell proliferation and cytotoxicity. By measuring the fluorescence intensity, cell growth and survival can be evaluated.