ChromoTek's Fluorescent 2-Hybrid (F2H)-Assay is an elegant assay to directly visualize and analyze Protein-Protein Interactions (PPIs) in living mammalian cells.

The rationale for the F2H-Assay is based on the observation that proteins are freely roaming the cell unless interactions with certain cellular components transiently immobilize and enrich them at specific structures.

We take advantage of a stable nuclear interaction platform, the protein-protein interaction biosensor (PPIB). A fluorescently labeled bait protein is tethered to the interaction platform and assayed for co-localization of a fluorescent prey fusion protein. The distinct read-out of the F2H-Assay is based on the optical co-localization of two distinguishable fluorophores. The respective dual flurorescent bright merge signal is clearly detectable and measurable under the microscope. Disruption of the binding by the test substances recreates a single flurorescent spot.

The F2H-Assay enables you to analyze mutation and deletion constructs and to map protein-protein interaction domains, in real time. The technology furthermore allows you to successfully investigate a multitude of interactions of proteins from different subcellular compartments and to perform library screens.

For drug discovery campaigns the F2H platform can be fully automated. One particular advantage of the F2H platform is that only those molecules will emege as possible drug candidates which are taken up into cells and which show little or no cytotoxicity. In addition, due to the standardized binding paltform used, it is possible to conduct comparative PPI studies.