5-Bromo-4-chloro-3-indolyl phosphate disodium salt, 5 g
Molar mass (M) 397,46 g/mol
Storage temp. -20 °C
Transport temp. ambient temp.
CAS No. [102185-33-1]
Highly soluble in water (20 mg/ml), poorly soluble in DMF.
Protect from light and humidity.
excl. VAT. | 5 g per Pack Qty.
Art. No. A155.4
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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.
|Application||Western, in situ, Histochemistry|
|Signal product||Precipitate (Blot)|
|Use||Western, in situ, histochimie|
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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.
|Appearance||almost white powder|
|Assay (titr.)||≥99,0 %|
|Water (KF)||5,8-7,8 %|