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Fluoppi


Fluoppi Red

  •  Red (Monti-Red)  joins the lineup of Fluoppi, a novel tool which allows you to visualize protein-protein interactions in living cells.
  • In combination with  Green (Azami Green) , it enables multicolor imaging!
mTOR-FKBP12 interaction

Visualization of mTOR-FKBP12 interaction by Fluoppi and Floppy Red

Protein protein interaction (PPI) of mTOR FRB domain and FKBP12 (FK506 binding protein) can be induced by Rapamycin. In this experiment, FKBP12 was fused to Ash-tag (FKBP12-Ash) and mTOR was fused to Monti-Red (mTOR-MR) or Azami-Green (mTORAG). Each color of mTOR construct was co-expressed with FKBP12-Ash then Rapamycin was added to the medium.

Screening of PPI (Protein-Protein Interaction) inhibitors

One of the outstanding features of Fluoppi is its reversibility. By adding PPI inhibitors, the pre-formed fluorescent foci would be dissociated in real time manner.This feature is perfectly suited for screening of PPI inhibitors in each phase of drug discovery program.

Inhibition of p53-MDM2 interaction
Inhibition of p53-MDM2interaction visualized by Fluoppi Red

Using p53-Ash-tag/MDM2-Monti-Red, Nutlin-3, a p53/MDM2 PPI inhibitor, was added to the medium after confirming the formation of Foci. Dissociation of Foci were observed in 10 minutes after the addition of Nutlin-3.

Multi-color Fluoppi: Visualizing dual PPIs in a single cell

mTOR (FRB domain) and FKBP12 is a well known PPI whose interaction is induced by Rapamycin. On the other hand, mTOR (FRB domain) and mCAB (the BBH region of Calcineurin A (residues 340-394) fused to Calcineurin B*) interaction is induced by FK506. In this experiment, The mCAB-hAG, mTOR-Monti-Red and Ash-FKBP12 were cotransfected into the cells at the same times. When the cells were exposed to Rapamycin, only the red foci were formed as shown in the left 4 pictures. On the other hand, in the case of FK506, only the green foci were detected. The results demonstrate the possibility that multi-color Fluoppi could be applied to visualize branched signaling pathway.
* Reference:P. A. Clemons et al. Chem. Biol. (2002)

Visualization of dual PPIs by Floppy and Fluoppi Red in a single cell

The new fluorescent protein, Monti-Red

Characteristics of  Monti-RedMonti-Red is a new red fluorescent protein that is developed for Fluoppi technology.
Among the many red fluorescent proteins, Monti-Red is relatively bright, allows clear observation of Foci under a fluorescence microscope, and is ideal for use in long-term imaging.

Monti-red spectrum

Oligomerization
Excit./Emiss. Maxima (nm)
Molar Extinction
Coefficient (M-1cm-1)
Fluorescence
Quantum Yield
Brightness
pH sensitivity
Cytotoxicity
Tetramer
571 / 607
83000
0.3
24.9
pKa=5.5
No







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