Enzyme-linked immunosorbent assay for trkA tyrosine kinase activity

被引：42
作者：
Angeles, TS
Steffler, C
Bartlett, BA
Hudkins, RL
Stephens, RM
Kaplan, DR
Dionne, CA
机构：
[1] NCI,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,EUKARYOT SIGNAL TRANSDUCT SECT,FREDERICK,MD 21702
[2] CEPHALON INC,DEPT CHEM,W CHESTER,PA 19380
来源：
ANALYTICAL BIOCHEMISTRY | 1996年 / 236卷 / 01期
关键词：
D O I：
10.1006/abio.1996.0130
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
A 96-well microtiter enzyme-linked immunosorbent assay was developed to assay the activity of the cytoplasmic domain of trkA tyrosine kinase. The assay involves immobilization of phospholipase C-gamma/glutathione S-transferase fusion protein on a microtiter plate, addition of the kinase reaction mixture, and detection by an antibody to phosphotyrosine followed by an alkaline phosphatase-conjugated second antibody. The substrate used in this system, phospholipase C-gamma, is one of several biologically important substrates for the phosphorylation reaction of receptor-linked tyrosine kinases. The assay was then used to characterize kinase inhibitory activities of various small molecules including analogs of K-252a. (C) 1996 Academic Press, Inc.
引用
收藏
页码：49 / 55
页数：7
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