Multiple transcription factor profiling by enzyme-linked immunoassay

被引：65

作者：

Shen, Z ^{[1
]}

Peedikayil, J ^{[1
]}

Olson, GK ^{[1
]}

Siebert, PD ^{[1
]}

Fang, Y ^{[1
]}

机构：

[1] BD Biosci Clontech, Palo Alto, CA 94303 USA

来源：

BIOTECHNIQUES | 2002年 / 32卷 / 05期

关键词：

D O I：

10.2144/02325dd07

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Transcription factor-DNA interactions have been widely studied in the regulation of gene expression. We have established an enzyme-linked immunoassay platform to quantify specific transcription factor-DNA interactions. In our assay; dsDNA immobilized on a 96-well plate captures the transcription factor from the nuclear extract of mammalian cells. The DNA-bound transcription factor is detected and quantified by enzyme-linked immunoassay using a transcription factor-specific antibody. We have profiled multiple transcription factors involved in inflammation including NFkappaB p50, NFkappaB p65, c-Rel, c-Fos, CREB-1, and ATF-2. When compared with the traditional electrophoretic mobility shift assay, the enzyme-linked immunoassay shows a 10 fold higher sensitivity, eliminates the use of radioactivity, allows for a high-throughput format, and is faster.

引用

页码：1168 / +

页数：7

共 17 条

[1] Nonisotopic quantitative analysis of protein-DNA interactions at equilibrium [J].