| Size | Price | Stock | Qty |
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| 5g |
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| Other Sizes |
| Targets |
As a substrate, pNPP does not have a biological target. It is a chromogenic substrate that is hydrolyzed by alkaline and acid phosphatases. The enzyme catalyzes the hydrolysis of pNPP to para-nitrophenol and inorganic phosphate. The product, para-nitrophenol, is yellow in alkaline conditions and can be quantified spectrophotometrically. This reaction is the basis for measuring phosphatase activity in biochemical and immunological assays.
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| ln Vitro |
The hydrolysis of PNPP is catalyzed by phosphatase, which releases inorganic phosphate and p-nitrophenol's conjugate base (pNP). The resultant phenolate has a maximum absorption wavelength of 405 nm and is yellow in color[1].
As a substrate, pNPP does not have direct pharmacological activity. Its activity is assessed by the rate of hydrolysis by phosphatases. In ELISA and other assays, the amount of para-nitrophenol produced is proportional to the amount of enzyme present. The compound's utility in vitro is its ability to serve as a sensitive and convenient substrate for detecting phosphatase activity. |
| ln Vivo |
pNPP is not used for in vivo pharmacological applications. It is an in vitro reagent for biochemical assays. No in vivo activity or therapeutic effects are associated with this compound. It is used exclusively in laboratory research and diagnostic applications.
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| Enzyme Assay |
Phosphatase activity assays using pNPP are performed in microplate or cuvette format. The substrate is dissolved in an appropriate buffer (e.g., 1 M diethanolamine buffer, pH 9.8, for alkaline phosphatase). Enzyme samples (e.g., alkaline phosphatase-conjugated antibodies in ELISA) are incubated with pNPP at 37°C for 15-60 minutes. The reaction is stopped by adding NaOH or other stop solution. Absorbance is measured at 405 nm using a spectrophotometer or microplate reader. The amount of para-nitrophenol produced is calculated using a standard curve. Each assay includes appropriate controls and blanks.
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| Cell Assay |
pNPP is not used in cell-based assays for pharmacological evaluation. It is a biochemical reagent used in ELISA and other enzyme activity assays. In these assays, the substrate is added to the reaction mixture, and the enzyme activity is measured by the production of the colored product. No cell culture protocols are applicable for this compound as a substrate.
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| Animal Protocol |
pNPP is not used in animal studies. It is an in vitro reagent for biochemical and diagnostic applications. No in vivo studies are conducted with this compound as it is not a drug candidate. It is used exclusively in laboratory research.
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| ADME/Pharmacokinetics |
pNPP has a molecular weight of 263.05 g/mol and a molecular formula of C6H4NNa2O6P. It is a white to off-white crystalline powder. Solubility: soluble in water. Storage: typically at room temperature, protected from light. As a biochemical reagent, it is stable under recommended storage conditions. No pharmacokinetic data are available as it is not a drug candidate.
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| Toxicity/Toxicokinetics |
pNPP is considered safe for laboratory use. It is not a drug candidate and has no therapeutic applications. Standard safety precautions for handling chemicals should be followed. It may cause irritation to eyes, skin, and respiratory tract. The compound is intended for research and diagnostic use only.
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| References | |
| Additional Infomation |
pNPP is also known as 4-Nitrophenyl phosphate disodium salt, PNPP disodium, and BNPP sodium salt. It is a non-proteinaceous chromogenic substrate for alkaline and acid phosphatases. It is widely used in ELISA and conventional spectrophotometric assays for measuring phosphatase activity. No clinical trials or regulatory approvals have been reported. The compound is for research and diagnostic use only.
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| Molecular Formula |
C6H4NNA2O6P
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|---|---|
| Molecular Weight |
263.0524
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| Exact Mass |
218.993
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| CAS # |
4264-83-9
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| Related CAS # |
330-13-2 (Parent)
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| PubChem CID |
77949
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| Appearance |
White to yellow solid powder
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| Density |
1.7±0.1 g/cm3
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| Boiling Point |
457.8±47.0 °C at 760 mmHg
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| Melting Point |
>300 °C(lit.)
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| Flash Point |
230.7±29.3 °C
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| Vapour Pressure |
0.0±1.2 mmHg at 25°C
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| Index of Refraction |
1.617
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| LogP |
0.37
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
16
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| Complexity |
238
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
VIYFPAMJCJLZKD-UHFFFAOYSA-L
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| InChi Code |
InChI=1S/C6H6NO6P.2Na/c8-7(9)5-1-3-6(4-2-5)13-14(10,11)12;;/h1-4H,(H2,10,11,12);;/q;2*+1/p-2
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| Chemical Name |
disodium;(4-nitrophenyl) phosphate
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O : ~160 mg/mL (~608.25 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (380.16 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.8016 mL | 19.0078 mL | 38.0156 mL | |
| 5 mM | 0.7603 mL | 3.8016 mL | 7.6031 mL | |
| 10 mM | 0.3802 mL | 1.9008 mL | 3.8016 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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