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5-Bromo-4-chloro-3-indolyl phosphate disodium salt, 100 mg

≥99 %
Pack Qty.
Pack.
BCIP-Na2, X-Phosphate disodium salt, X-Phos disodium salt
Empirical formula C8H4BrClNO4PNa2 · 1,5 H2O
Molar mass (M) 397,46 g/mol
Storage temp. -20 °C
Transport temp. ambient temp.
CAS No. [102185-33-1]

Substrate for alkaline phosphatase, in particular in immunoblot procedures.
Highly soluble in water (20 mg/ml), poorly soluble in DMF.

Protect from light and humidity.

In combination with NBT ideal for colorimetric detection of alkaline phosphatase activity, in partic ...
Product details
Type analysis

€28.95/Pack Qty. 

excl. VAT. | 100 mg per Pack Qty.

Art. No. A155.1

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5-Bromo-4-chloro-3-indolyl phosphate disodium salt ≥99 %

In combination with NBT ideal for colorimetric detection of alkaline phosphatase activity, in particular in immunoblot procedures.



Technical Information
Application Western, in situ, Histochemistry  
Enzyme Alkalische Phosphatase  
Detection Farbe (braun-violett)  
Signal product Präzipitat (Blot)  
Use Western, in situ, histochimie  
5-Bromo-4-chloro-3-indolyl phosphate disodium salt
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Order No. Pack Qty. Pack. Price Quantity
A155.1 100 mg glass €28.95
A155.2 500 mg glass €123.10
A155.3 1 g glass €211.70
A155.4 5 g glass €686.95
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General information

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.


Analysis certificates

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The following analysis certificates have been found:

Type analysis

Appearancealmost white powder  
Assay (titr.)≥99,0 %  
Water (KF)5,8-7,8 %  

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