4-Nitrophenyl phosphate  (Synonyms: p-Nitrophenyl phosphate)

4-Nitrophenyl phosphate (p-Nitrophenyl phosphate) is widely used as a small molecule phosphotyrosine-like substrate in activity assays for protein tyrosine phosphatases.4-Nitrophenyl phosphate is a colorless substrate that upon hydrolysis is converted to a yellow 4-nitrophenolate ion that can be monitored by absorbance at 405 nm^[1].

1. Equilibrate 4-Nitrophenyl phosphate to room temperature before opening.
2. Dissolve 10 mg of 4-Nitrophenyl phosphate in 10 mL of 1X Diethanolamine (HY-W035903) Substrate Buffer.
Note: Diethanolamine is effective at concentrations from 10 mM to 1 M.
3. Add 100 μL of the 4-Nitrophenyl phosphate solution to each microplate well.
4. Incubate plate at room temperature for 30 minutes, or until sufficient color develops.
5. Add 50 μL of 2 N NaOH to each well to stop the reaction.
6. Measure the absorbance at 405 nm. Alternatively, monitor reaction in a kinetic ELISA reader.

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

219.09

C₆H₆NO₆P

330-13-2

Solid

White to off-white

O=P(O)(O)OC(C=C1)=CC=C1[N+]([O-])=O

Room temperature in continental US; may vary elsewhere.

• [1]. Lountos GT, et al. Structural analysis of human dual-specificity phosphatase 22 complexed with a phosphotyrosine-like substrate. Acta Crystallogr F Struct Biol Commun. 2015 Feb;71(Pt 2):199-205.  [Content Brief]

  [2]. Qi S , et al. Development of a facile and sensitive method for detecting alkaline phosphatase activity in serum with fluorescent gold nanoclusters based on the inner filter effect. Analyst. 2020 Jun 7;145(11):3871-3877.  [Content Brief]