Figure 1

Experimental design of a microsphere-based immunoassay for measuring the activity of transcription factors ( MIA-TF ). a Flowchart of MIA-TF. The biotin-labeled dsDNA probes were made to react with TFs in a nuclear extract of the cells. Subsequently, NeutrAvidin microspheres were added into the reactions. After incubation, the mixtures were probed with anti-transcription factor antibody and detected with a fluorescent secondary antibody by flow cytometry. b Dot plot (SSC vs. FSC) of the MIA-TF method. The DNA-bound TFs were detected by an immunoassay using a transcription factor-specific antibody and then analyzed using flow cytometry. Microspheres not incubated with nuclear extract were used as negative controls (left) to gate the range of effective signals (R1). The microspheres incubated with the nuclear extracts increased the fluorescence values in the range of effective signals (right). bio-DNA biotin-labeled dsDNA, TF transcription factor.