Signosis’ TF-TF Interaction Plate Array can simultaneously profile the transcriptional interaction of multiple TFs with a TF or co-regulator of interest. In this assay, a series of unique biotin-labeled probes are provided that correspond with the consensus sequences of individual TF DNA-binding sites. Therefore, each probe represents an individual TF.

When the probe mix is incubated with nuclear extract, individual probes bind their corresponding TF. The TF or co-regulator of interest is then immunoprecipitated, along with transcriptionally interacting TFs, using a corresponding antibody and protein G or A agarose beads in a tube. Unbound probes and proteins are washed away. The bound probes are then detached from the complex and are subsequently denatured. The biotin-labeled DNA strands are hybridized on a pre-coat plate and detected with streptavidin-HRP and substrate. The detected signals reflect the interacting TFs with the particular TF or co-regulator. Luminescence is reported as relative light units (RLUs) on a microplate luminometer.