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PubMed · 12019791

Multiple transcription factor profiling by enzyme-linked immunoassay.

Z Shen·J Peedikayil·G K Olson·P D Siebert·Y Fang
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Abstract

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 transcriptionfactor-DNA interactions. In our assay, dsDNA immobilized on a 96-well plate captures the transcriptionfactor 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.

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Activating Transcription Factor 2AnimalsAntibody SpecificityBase SequenceCell ExtractsCell NucleusConsensus SequenceCyclic AMP Response Element-Binding ProteinDNAHeLa CellsHumansImmunoenzyme TechniquesJurkat CellsMammalsNF-kappa BNF-kappa B p50 SubunitProto-Oncogene Proteins c-fosSensitivity and SpecificityTranscription FactorsJournal Article

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Z Shen, J Peedikayil, G K Olson, P D Siebert, Y Fang. 2002. Multiple transcription factor profiling by enzyme-linked immunoassay.. https://doi.org/10.2144/02325dd07

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