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Detection of Alkaline Phosphatase Activity After Conventional Isoelectric Focusing by an Indoxyl-Tetrazolium Salt Technique

Detection of Alkaline Phosphatase Activity After Conventional Isoelectric Focusing by an Indoxyl-Tetrazolium Salt Technique

P Sinha

J Biochem Biophys Methods. 1985 Nov;11(6):327-40.

PMID: 2415567

Abstract:

Alkaline phosphatase was solubilized from human and rat tissues using papain in the presence of TRITON X-100 and subjected to isoelectric focusing (IEF) in polyacrylamide or agarose gels. Up till now, usually 1- and 2-naphthylphosphates have been used as substrates in order to specifically stain molecular forms of this enzyme by the azo-dye technique. In this paper, the use of another histochemical substrate, 5-bromo-4-chloro-3-indoxyl phosphate, in combination with tetrazolium salts [McGadey, J. (1970) Histochemie 23, 180-184] is presented. After hydrolysis, the released indoxyl moieties reduce tetrazolium salts to insoluble formazans at the zones of alkaline phosphatase activity. Zymogrammes showing molecular forms of alkaline phosphatase from 20 rat organs and the application of this staining technique for the detection of alkaline phosphatase activity in non-dialyzed human plasma after IEF are presented.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP3318432	Indoxyl phosphate disodium salt		3318-43-2	Price

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