Principle 

Signosis’ TF-TF Interaction Plate Array enables simultaneous profiling of multiple transcription factors (TFs) that interact with a specific TF or co-regulator of interest. The assay uses a mix of unique biotin-labeled DNA probes, each representing a consensus binding sequence for an individual TF.

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When incubated with nuclear extracts, the probes bind to their corresponding TFs. The TF or co-regulator of interest is then immunoprecipitated—along with any interacting TFs—using a specific antibody and Protein G or A agarose beads. After washing away unbound probes and proteins, the bound DNA probes are eluted, denatured, and hybridized to a pre-coated plate.

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Detection is performed using streptavidin-HRP and a chemiluminescent substrate. Luminescence is measured as relative light units (RLUs) using a microplate luminometer, providing insight into TF-TF interaction dynamics.