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: | , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2002-05-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/02325dd07 |
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| _version_ | 1850152171257987072 |
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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 |
| institution | OA Journals |
| 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 |
| work_keys_str_mv | AT zshen multipletranscriptionfactorprofilingbyenzymelinkedimmunoassay AT jpeedikayil multipletranscriptionfactorprofilingbyenzymelinkedimmunoassay AT gkolson multipletranscriptionfactorprofilingbyenzymelinkedimmunoassay AT pdsiebert multipletranscriptionfactorprofilingbyenzymelinkedimmunoassay AT yfang multipletranscriptionfactorprofilingbyenzymelinkedimmunoassay |