Multiple Transcription Factor Profiling by Enzyme-Linked Immunoassay

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 fac...

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Main Authors: Z. Shen, J. Peedikayil, G.K. Olson, P.D. Siebert, Y. Fang
Format: Article
Language:English
Published: Taylor & Francis Group 2002-05-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/02325dd07
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author Z. Shen
J. Peedikayil
G.K. Olson
P.D. Siebert
Y. Fang
author_facet Z. Shen
J. Peedikayil
G.K. Olson
P.D. Siebert
Y. Fang
author_sort Z. Shen
collection DOAJ
description 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 NFκB p50, NFκB 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.
format Article
id doaj-art-c9b89ff261d34cecb9509ebd6756cc99
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issn 0736-6205
1940-9818
language English
publishDate 2002-05-01
publisher Taylor & Francis Group
record_format Article
series BioTechniques
spelling doaj-art-c9b89ff261d34cecb9509ebd6756cc992025-08-20T02:26:03ZengTaylor & Francis GroupBioTechniques0736-62051940-98182002-05-013251168117710.2144/02325dd07Multiple Transcription Factor Profiling by Enzyme-Linked ImmunoassayZ. Shen0J. Peedikayil1G.K. Olson2P.D. Siebert3Y. Fang41BD Biosciences Clontech, Palo Alto, CA, USA1BD Biosciences Clontech, Palo Alto, CA, USA1BD Biosciences Clontech, Palo Alto, CA, USA1BD Biosciences Clontech, Palo Alto, CA, USA1BD Biosciences Clontech, Palo Alto, CA, USATranscription 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 NFκB p50, NFκB 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.https://www.future-science.com/doi/10.2144/02325dd07
spellingShingle Z. Shen
J. Peedikayil
G.K. Olson
P.D. Siebert
Y. Fang
Multiple Transcription Factor Profiling by Enzyme-Linked Immunoassay
BioTechniques
title Multiple Transcription Factor Profiling by Enzyme-Linked Immunoassay
title_full Multiple Transcription Factor Profiling by Enzyme-Linked Immunoassay
title_fullStr Multiple Transcription Factor Profiling by Enzyme-Linked Immunoassay
title_full_unstemmed Multiple Transcription Factor Profiling by Enzyme-Linked Immunoassay
title_short Multiple Transcription Factor Profiling by Enzyme-Linked Immunoassay
title_sort multiple transcription factor profiling by enzyme linked immunoassay
url https://www.future-science.com/doi/10.2144/02325dd07
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AT gkolson multipletranscriptionfactorprofilingbyenzymelinkedimmunoassay
AT pdsiebert multipletranscriptionfactorprofilingbyenzymelinkedimmunoassay
AT yfang multipletranscriptionfactorprofilingbyenzymelinkedimmunoassay