A MALACHITE GREEN COLORIMETRIC ASSAY FOR PROTEIN PHOSPHATASE-ACTIVITY

被引：329

作者：

GELADOPOULOS, TP ^{[1
]}

SOTIROUDIS, TG ^{[1
]}

EVANGELOPOULOS, AE ^{[1
]}

机构：

[1] NATL HELLENIC RES FDN, INST BIOL RES, 48 VASSILEOS CONSTANTINOU AVE, GR-11635 ATHENS, GREECE

来源：

ANALYTICAL BIOCHEMISTRY | 1991年 / 192卷 / 01期

关键词：

D O I：

10.1016/0003-2697(91)90194-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A simple and sensitive colorimetric assay for protein phosphatase activity based on the determination of released Pi by an improved malachite green procedure (A. A. Baykov, O. A. Evtushenko, and S. M. Avaeva, 1988, Anal. Biochem. 171, 266-270) is described. Proteins must be removed or stabilized prior to Pi determination with 0.25 n sulfuric acid or 3% (w/w) perchloric acid. Alternatively, to avoid possible acid hydrolysis of phosphate groups from organic compounds during deproteinization, the protein present in the phosphatase assay mixture can be stabilized with sodium dodecyl sulfate. In this case, the excess detergent is subsequently removed by precipitation with KCl because it colors with the malachite green reagent. The above procedure was applied to the determination of phosphorylase phosphatase activity in bovine brain extracts and the results are comparable to those obtained with the radioisotopic phosphatase assay. © 1991.

引用

页码：112 / 116

页数：5

共 21 条

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