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  3. 4-MUNANA

4-MUNANA is a substrate of influenza virus neuraminidase (NA) with high selectivity and irreversible reaction. In the enzymatic reaction, 4-MUNANA is hydrolyzed by NA to generate fluorescent 4-methylumbelliferone (4-MU). By detecting the fluorescence intensity of 4-MU, quantitative analysis of NA activity can be achieved. 4-MUNANA can be used in influenza-related research, such as screening NA inhibitors, developing new anti-influenza drugs, and studying the infection mechanism of influenza viruses.

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4-MUNANA Chemical Structure

4-MUNANA Chemical Structure

CAS No. : 76204-02-9

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Description

4-MUNANA is a substrate of influenza virus neuraminidase (NA) with high selectivity and irreversible reaction. In the enzymatic reaction, 4-MUNANA is hydrolyzed by NA to generate fluorescent 4-methylumbelliferone (4-MU). By detecting the fluorescence intensity of 4-MU, quantitative analysis of NA activity can be achieved. 4-MUNANA can be used in influenza-related research, such as screening NA inhibitors, developing new anti-influenza drugs, and studying the infection mechanism of influenza viruses[1][2].

In Vitro

In the experiment of screening neuraminidase inhibitors of traditional Chinese medicine, 4-MUNANA (50 μg/mL; react with NA for 30 minutes) can be used to evaluate the activity of NA by detecting the fluorescence intensity of 4-Methylumbelliferone (4-M) generated by its reaction with NA, thereby screening out traditional Chinese medicines such as Bupleurum Root that have inhibitory effects on NA, and determining their active ingredients and IC50[1].
The specific steps are as follows:
First, prepare a 50 μg/mL 4-MUNANA stock solution in PBS. Immerse the test strip in the stock solution for 30 seconds, then take it out and react with different concentrations of neuraminidase (NA) for 30 minutes.
After the reaction is completed, observe the fluorescence changes of the test strip under 365 nm light and record the image with a smartphone. At the same time, different concentrations of NA were added to the 4-MUNANA solution and allowed to react for 30 minutes. The fluorescence intensity of the 4-MU generated by the reaction was then measured (Ex=365 nm, Em=445 nm). This was used to establish a standard curve of NA concentration and fluorescence value for evaluating NA activity.
When screening NA inhibitors in Chinese herbal extracts, the Chinese herbal extracts were first reacted with NA, and then 4-MUNANA was added. The inhibitory effect of the extracts on NA was determined by detecting changes in fluorescence intensity.
4-MUNANA (0.025 mM; reacted with NA for 10 minutes) was used to measure the enzyme activity of NA in an experiment to study the functional balance between HA and NA of influenza viruses. The results showed that there was a functional balance between the activities of HA and NA in human pandemic viruses, while swine-derived viruses often deviated from this balance[2].
The specific steps are as follows:
The reaction was carried out in a reaction buffer containing 100mM imidazole-malic acid (pH 6.15),150mM NaCl, 10mM CaCl2, 0.1mg/mL bovine serum albumin and 0.02% NaN3. 4-MUNANA was used as a substrate, and its concentration was 0.025 mM when used in the experiment to compare the enzyme activity under different NA concentrations. The total reaction volume was 80μL, and the reaction was carried out at room temperature (22°C) for 10 minutes, followed by the addition of 80μl 1M Na2CO3 to terminate the reaction. Finally, the concentration of the reaction product was measured (Ex=365 nm, Em=450 nm) to determine the enzyme activity of NA and study the relationship between the functional balance of HA and NA in influenza virus and virus transmission.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

489.41

Formula

C21H24NNaO11

CAS No.
Appearance

Solid

Color

White to light yellow

Emission (Em)

410

Excitation (Ex)

317

SMILES

O=C(O[Na])[C@@]1(O[C@@]([C@@H]([C@H](C1)O)NC(C)=O)([H])[C@H](O)[C@H](O)CO)OC2=CC=C(C(C)=CC(O3)=O)C3=C2

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

-20°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

Solvent & Solubility
In Vitro: 

H2O : 233.33 mg/mL (476.76 mM; Need ultrasonic)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.0433 mL 10.2164 mL 20.4328 mL
5 mM 0.4087 mL 2.0433 mL 4.0866 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Working solution concentration: mg/mL
This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact MedChemExpress (MCE).
Purity & Documentation

Purity: 98.08%

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
H2O 1 mM 2.0433 mL 10.2164 mL 20.4328 mL 51.0819 mL
5 mM 0.4087 mL 2.0433 mL 4.0866 mL 10.2164 mL
10 mM 0.2043 mL 1.0216 mL 2.0433 mL 5.1082 mL
15 mM 0.1362 mL 0.6811 mL 1.3622 mL 3.4055 mL
20 mM 0.1022 mL 0.5108 mL 1.0216 mL 2.5541 mL
25 mM 0.0817 mL 0.4087 mL 0.8173 mL 2.0433 mL
30 mM 0.0681 mL 0.3405 mL 0.6811 mL 1.7027 mL
40 mM 0.0511 mL 0.2554 mL 0.5108 mL 1.2770 mL
50 mM 0.0409 mL 0.2043 mL 0.4087 mL 1.0216 mL
60 mM 0.0341 mL 0.1703 mL 0.3405 mL 0.8514 mL
80 mM 0.0255 mL 0.1277 mL 0.2554 mL 0.6385 mL
100 mM 0.0204 mL 0.1022 mL 0.2043 mL 0.5108 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Product Name:
4-MUNANA
Cat. No.:
HY-W019823
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