PNPP

SKU: orb1307864

Description

PNPP (p-Nitrophenyl Phosphate) is a chromogenic substrate for detecting alkaline and acid phosphatase activity. It is widely used in ELISA and spectrophotometric assays for in vitro research, including enzyme kinetics and diagnostic applications in clinical and biochemical studies.

Key Properties

• CAS Number: 4264-83-9
• MW: 263.05
• Purity: 99.83%
• Formula: C₆H₄NNa₂O₆P
• SMILES: [Na+].[Na+].[O-][N+](=O)c1ccc(OP([O-])([O-])=O)cc1
• Target: Others
• Solubility: H2O:255 mg/mL (969.4 mM)

Bioactivity

• In Vitro:
  Instructions I. Solution preparation 1. Stock solution preparation: Dissolve PNPP powder in an appropriate buffer, usually at a concentration of 5–10 mM. It is recommended to use a buffer containing 50 mM Tris-HCl and 10 mM MgCl₂, with the pH adjusted to 9.5 to suit the activity of alkaline phosphatase. 2. Working solution preparation: Dilute the stock solution to the required concentration, usually 2–5 mM, according to experimental needs. II. Operation steps 1. Reaction conditions: Temperature: 37°C. Time: 10–30 minutes, the specific time needs to be optimized according to the experiment. 2. Detection method: 1) Under the action of the enzyme, PNPP is hydrolyzed to produce yellow p-nitrophenol, which has a maximum absorption peak at 405 nm. 2) Use a spectrophotometer to measure the absorbance at 405 nm to evaluate the activity of the enzyme. Note: Fresh preparation: PNPP solution should be prepared before use and avoid long-term storage to prevent hydrolysis. Store away from light: PNPP is light-sensitive, so strong light exposure should be avoided during operation. Termination of reaction: In ELISA experiments, an equal volume of 0.1 M NaOH or 0.1 M EDTA can be added after the reaction is completed to terminate the reaction.

Storage & Handling

• Storage: store at low temperature,keep away from direct sunlight | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice/Shipping at ambient temperature.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

PNPP

Compound Identifiers

PubChem CID

77949