F2H Kits p53-Mdm2/4

Description
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.

Specificity
Bait:    GFP-p53
Prey:   RFP-Mdm2 or RFP-Mdm4
Bait and prey interact.

Application
Interaction analysis between the tumor suppressor p53 and its negative regulators Mdm2 or Mdm4:
Monitor protein dynamics upon treatments
Screen protein interaction inhibitors
Click on left picture to activate images and animation

Product Size Code Price Buy
Product F2H®-Kit p53/Mdm2 Size F2H®-BHK Cell line: 5x106 cells, Platform-p53-Mdm2: 100 µg, Control-p53: 30 µg Code f2hk-p53/mdm2 Price $ 1385
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Product F2H®-Kit p53/Mdm4 Size F2H®-BHK Cell line: 5x106 cells, Platform-p53-Mdm4: 100 µg, Control-p53: 30 µg Code f2hk-p53/mdm4 Price $ 1385
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Product Platform-p53-Mdm2 Size 100 µg Code f2h-ppm2 Price $ 525
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Product Platform-p53-Mdm4 Size 100 µg Code f2h-ppm4 Price $ 525
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Product Control-p53 Size 30 µg Code f2h-cp Price $ 120
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Cell line
BHK (baby hamster kidney)

Detection
Epifluorescence microscope

  • 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)

Assay principle

  • 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.

Results

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

Click on picture at the top of the page to see all animations.

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

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