AUTOMATED-ASSAY OF PHOSPHOHYDROLASES BY MEASURING THE RELEASED PHOSPHATE WITHOUT DEPROTEINIZATION

被引：45

作者：

HEGYVARY, C

KANG, K

BANDI, Z

机构：

[1] RUSH UNIV,DEPT PHYSIOL,CHICAGO,IL 60612

[2] RUSH UNIV,DEPT BIOCHEM,CHICAGO,IL 60612

来源：

ANALYTICAL BIOCHEMISTRY | 1979年 / 94卷 / 02期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1016/0003-2697(79)90380-4

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe an automated determination of inorganic phosphate in the presence of proteins and its application for the assay of NaK-ATPase (EC 3.6.1.3) and other insoluble phosphohydrolases. After incubation the enzyme reaction is stopped at neutral pH with 3.3% (w/v) sodium dodecyl sulfate plus 30 mm EDTA (final concentration). The released phosphate is measured on the Technicon Autoanalyzer as phosphomolybdate reduced with ferrous ammonium sulfate and thiourea to molybdenum blue. EDTA enhances color development and solubilization of the proteins. The reagents are stable at room temperature and are formulated from cheap, common chemicals. © 1979.

引用

页码：397 / 401

页数：5

共 17 条

[11] PURIFICATION AND PROPERTIES OF A HIGHLY ACTIVE OUABAIN-SENSITIVE NAT K+-DEPENDENT ADENOSINETRIPHOSPHATASE FROM CARDIAC TISSUE [J].