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 |
| Published: |
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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| Summary: | 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. |
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| ISSN: | 0736-6205 1940-9818 |