F2H®-Kits p53-Mdm2/4

The Fluorescent Two-Hybrid (F2H®) Kits for p53-Mdm2 and for p53-Mdm4 respectively enable intracellular analysis of the interaction between the tumor suppressor p53 and its negative regulators Mdm2 or Mdm4.

  • GFP-tagged human p53 is anchored at a specific location in the nucleus of  F2H®-Cells, forming a bright fluorescent spot in the green channel.
  • Interaction with the RFP-tagged human Mdm2 or the RFP-tagged human Mdm4 can be easily evaluated by conventional fluorescence microscopy as enrichment of red fluorescence at the location of the green spot.
  • Compounds’ ability to disrupt the p53 and Mdm2 interaction or the p53 and Mdm4 interaction is determined based on the disappearance of the red spots from the anchored green spots.

F2H p53-Mdm4
  • Monitor protein dynamics upon treatments
  • Screen protein interaction inhibitors

Please inquire if you want to learn more about our F2H® Assay Kits.

Results

F2H® allows direct side-by-side comparison of the compound's activity and kinetics in live cells.

p53:Mdm2 + Nutlinp53:Mdm2 + sMTide02
p53:Mdm4 + Nutlinp53:Mdm4 + sMTide02 

Incubations with 50 μm Nutlin-3 or stapled peptide sMTide-02 were carried out for 7 h during imaging. Fading of the red fluorescence from the spot but not from the rest of the nucleus indicates disruption of the interaction.

Ordering
information

Prices in EUR

ProductCodePriceBuy
F2H®-Kit p53/Mdm2 
  • F2H®-BHK Cell Line (5x106 cells)
  • Platform-p53-Mdm2
  • Control-p53
 
f2hk-p53/mdm2990 EUR Cart
F2H®-Kit p53/Mdm4 
  • F2H®-BHK Cell Line (5x106 cells)
  • Platform-p53-Mdm4
  • Control-p53
f2hk-p53/mdm4990 EUR Cart
Platform-p53-Mdm2 (100µg)f2h-ppm2375 EURCart
Platform-p53-Mdm4 (100µg)f2h-ppm4375 EURCart
Control-p53 (30µg)f2h-cp85 EUR Cart

Prices in USD

ProductCodePriceBuy
F2H®-Kit p53/Mdm2 
  • F2H®-BHK Cell Line (5x106 cells)
  • Platform-p53-Mdm2
  • Control-p53
 
f2hk-p53/mdm21385 USD Cart
F2H®-Kit p53/Mdm4 
  • F2H®-BHK Cell Line (5x106 cells)
  • Platform-p53-Mdm4
  • Control-p53
f2hk-p53/mdm41385 USD Cart
Platform-p53-Mdm2 (100µg)f2h-ppm2525 USDCart
Platform-p53-Mdm4 (100µg)f2h-ppm4525 USDCart
Control-p53 (30 µg)f2h-cp120 USD Cart
Related
information
References

Latest References

  • L. Yurlova;M. Derks;A. Buchfellner;I. Hickson;M. Janssen;D. Morrison;I. Stansfield;C. J. Brown;F. J. Ghadessy;D. P. Lane;U. Rothbauer;K. Zolghadr;E. Krausz(2014). The Fluorescent Two-Hybrid Assay to Screen for Protein-Protein Interaction Inhibitors in Live Cells: Targeting the Interaction of p53 with Mdm2 and Mdm4 J Biomol Screen. (PubMed)
  • A. Wolf;M. Mantri;A. Heim;U. Muller;E. Fichter;M. M. Mackeen;L. Schermelleh;G. Dadie;H. Leonhardt;C. Venien-Bryan;B. M. Kessler;C. J. Schofield;A. Bottger(2013). The polyserine domain of the lysyl-5 hydroxylase Jmjd6 mediates subnuclear localization Biochem J. (PubMed)
  • S. J. Wei;T. Joseph;A. Y. Sim;L. Yurlova;K. Zolghadr;D. Lane;C. Verma;F. Ghadessy(2013). In vitro selection of mutant HDM2 resistant to Nutlin inhibition PLoS One. (PubMed)
  • C. J. Brown;S. T. Quah;J. Jong;A. M. Goh;P. C. Chiam;K. H. Khoo;M. L. Choong;M. A. Lee;L. Yurlova;K. Zolghadr;T. L. Joseph;C. S. Verma;D. P. Lane(2013). Stapled Peptides with Improved Potency and Specificity that Activate p53 ACS Chem Biol. (PubMed)
  • O. Mortusewicz;E. Fouquerel;J. C. Ame;H. Leonhardt;V. Schreiber(2011). PARG is recruited to DNA damage sites through poly(ADP-ribose)- and PCNA-dependent mechanisms Nucleic Acids Res. (PubMed)


see here for complete reference list.

Only for research applications, not for diagnostic or therapeutic use.