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5-Bromo-4-chloro-3-indolyl phosphate p-toluidine salt, 5 g

≥99 %, BioScience Grade
Examples of effect: Have an allergenic effect, carcinogenic, mutagenic, toxic to reproduction and cause developmental toxicity (reprotoxic) or damage to organs Safety: You must be well-informed before starting work with these substances; wear protective clothing and gloves, eye protection and mask or breathing protection.
i may cause damage to organs
P260 P280 P308+P311
i do not breathe dust, wear protective gloves/protective clothing, IF exposed or concerned: Call a POISON CENTER or doctor
Pack Qty.
BCIP-p-toluidine salt, X-Phos
Empirical formula C15H15N2O4BrClP
Molar mass (M) 433,63 g/mol
Melting point (mp) 198 °C
Storage temp. -20 °C
Transport temp. ambient temp.
CAS No. 6578-06-9
EG-Nr. 229-506-1

Histochemical substrate for phosphatase.
Highly soluble in DMF (20 mg/ml), poorly soluble in water.

Protect from light and humidity.

In combination with NBT ideal for colorimetric detection of alkaline phosphatase activity, in immunohistochemical procedures.
Product details

€338.65/Pack Qty. 

excl. VAT. | 5 g per Pack Qty.

Art. No. 6368.3

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Product details

5-Bromo-4-chloro-3-indolyl phosphate p-toluidine salt ≥99 %, BioScience Grade

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


By enzymatic dephosphorylation BCIP is converted to indoxyl which subsequently tautomerizes to the ketone. In basic pH values, this ketone dimerizes and, in doing so, releases H+ ions. The dye precipitates in the direct vicinity of the phosphatase, hence staining this site in a weak blue colour. Enhencement and darkening of this colour is achieved by adding NBT (Art. No. 4421).

Technical Information
Application Western, in situ, Histochemistry 
Enzyme Alkaline phosphatase 
Detection Colour (brown) 
Signal product Precipitate (Blot) 
Use Western, in situ, histochimie 
5-Bromo-4-chloro-3-indolyl phosphate p-toluidine salt
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6368.1 500 mg glass


6368.2 1 g glass


6368.3 5 g glass


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Downloads / MSDS

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.

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Guarantee analysis

Appearancewhite to almost white powder
Assay (titr.)≥99 %
Water (KF)≤1 %
Appearance of solution (1% in DMF)colourless to slightly yellowish, clear
Phosphate (PO4)≤0,05 %