Description F2H?-Kit Basic allows studying interactions between GFP- and RFP-tagged proteins in live mammalian cells.
Specificity Bait: GFP-fusion protein Prey: RFP-fusion protein, mCherry-fusion protein Tested on cytoplasmic, nuclear, and mitochondrial proteins Cell line: BHK (baby hamster kidney)
Application Visualize protein-protein interactions inside cells Validate your pull-down results or Y2H hits in mammalian cells Identify interacting protein domains
Principle
- F2H cells (BHK origin) carry a GFP-anchoring platform in the nucleus
- GFP- and RFP-fusion proteins (bait and prey) are transiently expressed in F2H cells
- GFP-bait is recruited to the platform → bright green spot
- If RFP-prey interacts with the bait → red fluorescent spot co-localizes with the green spot
- If RFP-prey does not interact with the bait → no red spot, disperse red fluorescence