F2H Kit Basic

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

Product Size Code Price Buy
Product F2H®-Kit Basic Size F2H®-BHK Cell line: 5x106 cells, Platform reagent: 100 µg, Control DNA: 30 µg Code f2hk Price $ 1230
Buy +
Product Platform reagent Size 100µg Code f2h-p100 Price $ 345
Buy +
Product Control DNA Size 30 µg Code f2h-c30 Price $ 120
Buy +

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)

 

 

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

F2H Kit basic assay 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

Benefits

See your proteins interact – live!

  • Visualize protein-protein interactions inside cells
  • Validate your pull-down results or Y2H hits in mammalian cells
  • Identify interacting protein domains

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

Experimental procedure

  • You get the F2H cell line
  • You get the Platform Reagent
  • You have your GFP-bait and RFP-prey plasmids

Day 1: Transfect the bait and the prey plasmids with the Platform Reagent into the F2H® cells

Day 2: Image and quantify your results

Hear from our customers

"I already had all flurescent protein-constructs in my fridge, therefore F2H was the fastest way to map the interaction domains."

Dr. Oliver Mortusewicz, Karolinska Institutet, Sweden

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