4-Methylumbelliferyl phosphate

4-Methylumbelliferyl phosphate (4-MUP), an anionic organophosphate, is a acid and alkaline phosphatase fluorogenic substrate. 4-Methylumbelliferyl phosphate is also a nerve agent simulant^[1][2][3]. In Vitro: Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Serum acid phosphatase assay^[1]:
1. Preparation a 100 μL solution containing the following agents: 5.0 μL of serum enzyme, 50 μL of 4-Methylumbelliferyl phosphate (5.0 mM), and 10 μL of 1.0 M buffer at pH 6.0.
2. Add the following agents (with the final concentration) 20 mM Sodium Tartrate (HY-128476), 15 mM Sodium Fluoride (HY-B1766), 3000 mM 2-Mercaptoethanol, 4 mM 1, 10-Phenanthroline (HY-W004544), 6 mM 8-Quinolinol (HY-B1005), 6 mM 8-Hydroxy-5quinolinesulfonic acid, 45 mM EDTA (HY-Y0682) and 6 mM EGTA (HY-D0861).
3. Incubate reaction mixtures for 15 min at 37℃ and terminated by the addition of 2.9 mL of 0.1 M ammonium hydroxide/glycine buffer, pH 10.5.
4. Mix samples thoroughly and determine fluorescence by a fluorometer.