PROTEIN PHOSPHATASE ASSAY SUITABLE FOR ACID-LABILE SUBSTRATES

被引：12

作者：

KIM, Y

MATTHEWS, HR

机构：

[1] Department of Biological Chemistry, School of Medicine, University of California, Davis

来源：

ANALYTICAL BIOCHEMISTRY | 1993年 / 211卷 / 01期

关键词：

D O I：

10.1006/abio.1993.1227

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A sensitive protein phosphatase assay suitable for acid-labile substrates has been developed. The assay involves incubation of the phosphatase with a 32P-labeled phosphoprotein substrate, quenching the reaction, and then filtering through an ultrafilter that retains substrate and allows the released phosphate to flow through. The released phosphate is counted. The use of a slightly alkaline quenching solution makes the assay suitable for all phosphoamino acids that are stable near physiological pH, including the acid-labile phosphoamino acids. The process is made very convenient by the use of disposable microcon-3 ultrafiltration devices. Radioactive waste is limited to reaction tubes, the microcon-3s, pipette tips, and the scintillation vials and fluid. © 1994 Academic Press, Inc. All rights reserved.

引用

页码：28 / 33

页数：6

共 19 条

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[2] PHOSPHORYLATION OF NUCLEAR PROTEINS IN RAT REGENERATING LIVER [J].