4-MUNANA is a fluorescent substrate employed in neuraminidase activity assays and serves as a neuraminidase substrate analogue.

Instructions:
I. Solution preparation
1. Stock solution: Dissolve 4-MUNANA in a suitable buffer to prepare a stock solution with a concentration of 1–10 mM. The specific concentration can be adjusted according to the experimental requirements.
2. Working solution: Dilute the 4-MUNANA stock solution to a working concentration (usually 100–500 µM). The specific concentration can be adjusted according to the experimental requirements.
Notes:
Powder: Store at -20°C or lower in a dark, dry environment.
Stock solution: Store at -20°C in a dark place after aliquoting. Avoid repeated freezing and thawing. Store at 4°C for a short period of time.
II. Operation steps
Neuraminidase activity detection
1. Detection buffer: Prepare a buffer suitable for neuraminidase activity, such as 50 mM sodium acetate (pH 5.5).
2. Enzyme reaction: Mix a sample containing neuraminidase (such as a virus sample or cell lysate) with the 4-MUNANA working solution.
3. Incubate at 37°C for 30–60 Minutes (time can be optimized according to the experiment).
4. After the reaction is completed, add an equal volume of stop solution (such as 0.1 M glycine buffer, pH 10.4) to enhance the fluorescence signal.
5. Fluorescence detection
Detection method:
Use a fluorescence spectrophotometer or microplate reader to detect the fluorescence signal generated by the reaction.
Excitation wavelength: ~365 nm
Emission wavelength: ~450 nm
Notes
Standard curve: Use known concentrations of 4-methylumbelliferone to prepare a standard curve for quantitative analysis of neuraminidase activity.
Control experiment: Set up negative control (no enzyme) and positive control (containing known inhibitors) to verify the specificity of the detection.

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

DMF: 15 mg/mL (30.65 mM), Sonication is recommended.

H2O: 80 mg/mL (163.46 mM), Sonication is recommended.

Ethanol: 0.25 mg/mL (0.51 mM), Sonication is recommended.

PBS (pH 7.2): 10 mg/mL (20.43 mM), Sonication is recommended.

DMSO: 255 mg/mL (521.04 mM), Sonication is recommended.