A SPECIFIC IMMUNOPRECIPITATION ASSAY FOR THE PROTEIN KINASE-F(A)/GLYCOGEN SYNTHASE KINASE-3

被引：27

作者：

VANLINT, J ^{[1
]}

KHANDELWAL, RL ^{[1
]}

MERLEVEDE, W ^{[1
]}

VANDENHEEDE, JR ^{[1
]}

机构：

[1] CATHOLIC UNIV LEUVEN,FAC GENEESKUNDE,AFDELING BIOCHEM,CAMPUS GASTHUISBERG,B-3000 LOUVAIN,BELGIUM

来源：

ANALYTICAL BIOCHEMISTRY | 1993年 / 208卷 / 01期

关键词：

D O I：

10.1006/abio.1993.1018

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A specific immunoprecipitation assay has been developed for the accurate determination of the kinase FA/GSK3 activity in crude cell preparations. The assay is based on the production and isolation of polyclonal peptide antibodies toward the C-terminus of the rat GSK3-α and GSK3-β isoforms. The respective forms of the kinases are captured as active immunocomplexes with immobilized secondary antibodies and their kinase activity toward the synthetic peptide P-GS1 can be accurately measured. Immunoprecipitation with a mixture of the two peptide antibodies allows for the detection and estimation of the total kinase FA/GSK3 activity in cellular extracts, whereas the α-peptide antibody specifically detects and measures the GSK3-α isoform. The contribution of GSK3-β in the total kinase FA/GSK3 activity of cell fractions can be calculated. © 1993 Academic Press, Inc.

引用

页码：132 / 137

页数：6

共 32 条

[1] THE ATP,MG-DEPENDENT PROTEIN PHOSPHATASE - REGULATION BY CASEIN KINASE-1 [J].