ROTI®Fair pNPP 20mg , 24 unit(s), blister
Transport temp. ambient temp.
0,1 % pNPP solution (1 mg/ml) is prepared by dissolution of 1 tablet in 20 ml substrate buffer.
excl. VAT. | 24 unit(s) per Pack Qty.
Art. No. 1128.1
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Exactly pre-weighed tablets for convenient and rapid preparation of pNPP solution.
Prepared solution may be sterilized by filtration (0,22 µm). The solution has to be stored at -20 °C in aliquots.
ROTI®Fair pNPP 20mg 20 mg/tablet, for biochemistry
|Batch quantity||20 mg/tablet|
|End concentration||1 mg/ml|
|Buffer/solution||pNPP (tablet), p-Nitrophenyl phosphate|
|Signal product||soluble (ELISA, activity assay)|
|Use||Substrate solution for alkaline phosphatase|
- Subtotal: 0.00
|Order No.||Pack Qty.||Pack.||Price||Quantity|
Available at short notice
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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.
|Prepared solution contains 1 mg/ml pNPP, when 1 tablet is dissolved in 20 ml substrate buffer or deionized, or distilled, water.|