Technical Data Sheet
4-Nitrophenyl phosphate disodium salt hexahydrate, 2.5 g
Molar mass (M) 371,12 g/mol
Melting point (mp) >300 °C
Storage temp. -20 °C
Transport temp. ambient temp.
WGK 1
CAS No. 4264-83-9
EG-Nr. 224-246-5
€19.90/Pack Qty.
excl. VAT. | 2.5 g per Pack Qty.
Art. No. 4165.1
Product details
4-Nitrophenyl phosphate disodium salt hexahydrate ≥99 %, for biochemistry
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.
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.
| Enzyme | Alkaline phosphatase |
| Detection | Colour (yellow) |
| Signal product | soluble (ELISA, activity assay) |
- Subtotal: 0.00
| Art. No. | Pack Qty. | Pack. | Price | Quantity | |
|---|---|---|---|---|---|
| 4165.1 | 2.5 g | glass |
€19.90 |
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| 4165.7 | 100 g | glass |
€235.45 |
|
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| 4165.9 | 25 g | glass |
€84.40 |
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In stock
Available
In procurement
No longer available
Delivery date currently unknown
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- Subtotal: 0.00
Downloads / MSDS
General information
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.
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.
Certificates of Analysis
Type analysis
| Appearance | off-white to light-yellow crystalline powder |
| Assay (HPLC) | ≥99.0 % |
| Water | 27.0-31.5 % |
| pH value (5 % in water) | 8.0-10.5 |