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4-Nitrophenyl phosphate disodium salt hexahydrate, 2.5 g

≥99 %, p.a.
p-Nitrophenyl phosphate, pNPP
Empirical formula C6H4NNa2O6P · 6 H2O
Molar mass (M) 371,12 g/mol
Melting point (mp) 300 °C
Storage temp. +4 °C
CAS No. [4264-83-9]
EG-Nr. 224-246-5

Substrate for alkaline phosphatase, particularly for ELISA.
Application: Working solution: 1 mg/ml in diethanolamine substrate buffer (10 mM diethanolamine, 0.5 mM MgCl2, pH 9.5). For endpoint analysis the reaction may be stopped via addition of 2–3 M NaOH.

Substrate of the alkaline phosphatase for use in ELISAs. During reaction of the substrate, a soluble ...
Guarantee analysis

16,05 €/VE 

Excl. btw | 2,5 g Per VE

Bestelnr. 4165.1

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4-Nitrophenyl phosphate disodium salt hexahydrate ≥99 %, p.a.

Substrate of the alkaline phosphatase for use in ELISAs. During reaction of the substrate, a soluble, yellow product is formed that is detected photometrically at 405 nm wave length.

Technische informatie
Enzyme Alkalische Phosphatase  
pKa (at 25 °C) ≥99 %  
Detection Farbe (gelb)  
Signal product löslich (ELISA, Aktivitätsassay)  
4-Nitrophenyl phosphate disodium salt hexahydrate
Geselecteerde hoeveelheid:   0
  1. Tussentotaal:  0.00
Bestelnr. Verp. Pack. Prijs Hoeveelheid
4165.1 2,5 g glass 16,05 €
4165.7 100 g glass 177,40 €
4165.9 25 g glass 65,05 €
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Geselecteerde hoeveelheid:   0
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Algemene informatie

Assay protocol for use of BCIP/NBT in immunoblot procedures:

Stock solutions (All solutions are stable for at least 1 year at 4 °C):
0.5 g NBT in 10 ml 70 % dimethylformamide;
0.5 g BCIP (p-toluidine salt) in 10 ml 100 % dimethylformamide.
Incubation buffer for alkaline phosphatase:
100 mM NaCl, 5 mM MgCl2,100 mM Tris (pH 9.5).
Fresh substrate solution:
66 μl NBT stock solution + 10 ml incubation buffer, mix well, add 33 μl BCIP stock solution. Use within 1 hour.
Blot development:
Approx. 10 ml substrate solution per 15 x 15 cm2 membrane surface. Develop at room temperature until bands become visible (approx. 30 mins).
Reaction stop: Rinse with PBS/20 mM EDTA.

Detection of peroxidase activity (Immunoassay):

Dissolve 1 mg TMB in 0.1 ml dimethylsulfoxide (Art. No. 4720); add 9.9 ml of a 0.1 M sodium acetate solution (pH 6.0) (Powder: Art. No. 6779), filter and add H2O2 (Art. No. 8070) (final concentration 0.01 %).

Always prepare freshly!

Incubation 10–30 mins at room temperature (approx. 50 μl per microtitre well; finally add 50 μl 1 M H2SO4, Art. No. X873, per well).
Photometric quantitation at 450 nm.

Reference: Bos, E.S. et al., (1981) J. Immunoassay 2, (3/4), 187.


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Guarantee analysis

Assay (HPLC)≥99,0 %
Solution (5 % in H2O)clear, colourless to yellow
Free 4-nitrophenol≤0,01 %

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