PNPP, Disodium Salt (CAS 4264-83-9)

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Synonym:	4-Nitrophenyl phosphate disodium salt solution
Application:	An alkaline phosphatase substrate used in ELISA applications; absorbs light at 405nm
CAS Number:	4264-83-9
Molecular Weight:	371.14
Molecular Formula:	C₆H₄NO₆P•2Na•6H₂O

Ordering Information

Product Name	Catalog #	Unit	Price	Qty	Add
PNPP, Disodium Salt	sc-3720	25 g	$115
PNPP, Disodium Salt	sc-3720A	250 g	$440

Description

A colorimetric alkaline phosphatase soluble substrate, p-Nitrophenyl Phosphate (pNPP) is the substrate of choice for use with alkaline phosphatase in Enzyme Linked Immunosorbant Assay (ELISA) procedures. The reaction produces a soluble end product that is yellow in color. Can be used to perform kinetic and endpoint determinations. Also used as a phosphorylating agent in nucleotide synthesis. Recommended for use with pNPP Substrate Buffer (sc-296099). Also available in solution form: p-Nitrophenyl Phosphate 50X solution (sc-358718).

Usage

• To prepare PNPP substrate solution, dissolve 10 mg of room-temperature PNPP, Disodium Salt (sc-3720) into 10 ml of pNPP Substrate Buffer (sc-296099). Mix well and protect from light. Diluted product may be used for up to 10 hours following mixing. Discard if solution becomes yellow or turbid.
• Following incubation of antibody samples and the addition of alkaline phosphatase labeled probes, wash plates thoroughly avoiding the use of phosphate buffers. Add 100 µl of 1X PNPP substrate solution to each well and incubate 15-60 minutes. Incubation times will vary depending on the enzyme present.
• For endpoint analysis, the reaction may be stopped with 3N NaOH and read at 405 nm.

Technical Information

Appearance:	White to yellow crystalline powder
Physical State:	Solid
Solubility:	Soluble in water(freely soluble). Insoluble in ethanol, and acetone.
Storage:	Store at -20° C
Melting Point:	>300 °C
Boiling Point:	457.8 °C at 760 mmHg (Predicted)

Safety and Reference Information

WGK Germany:	3
PubChem CID:	2723710
EC Number:	224-246-5
Beilstein Registry:	5324661
SMILES:	C1=CC(=CC=C1[N+](=O)[O-])OP(=O)([O-])[O-].O.O.O.O.O.O.[Na+].[Na+]

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.

References

1. Voller, A., et al. 1976. Bull. World Health Organ. 53: 55-65. PMID: 1085667
2. Engvall, E. 1980. Meth. Enzymol. 70: 419-439. PMID: 6999296
3. Voller, A., et al. 1986. Manual of Clinical Laboratory Immunology, 3rd Edition. In Rose, N., et al. eds., Washington, D.C. American Society for Microbiology, 106-107.

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