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One- and Two-Hybrid Systems

ACE01 - Grow'n'Glow ACE1 Two-Hybrid System

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The ACE1 Two-Hybrid System is a yeast two-hybrid system that efficiently identifies protein-protein interactions in vivo. Yeast two-hybrid systems depend on the ability of specific separated DNA binding and transactivation domains to reconstitute an active transcription factor complex when interacting proteins are fused to these domains. Clones that express interacting proteins are consequently detected through the activity of reporter genes.

The ACE1 Two-Hybrid System identifies protein-protein interactions by combining the advantages of the copper-inducible transcription factor ACE1 and the copper resistance-mediating CUP1 gene, with the proven green fluorescence protein (GFP) as the reporter. With this combination, the interactions of two proteins fused to the DNA-binding and transactivation domains of ACE1 can be detected by growth of the cells on copper-containing media and the copper-dependent expression of the strongly-fluorescent GFPuv, which causes those colonies to fluoresce bright green under UV light. The use of both a growth selection and a strong fluorescence signal creates a fast, highly sensitive and more reliable method of monitoring protein interactions in living cells. The copper concentration can be changed to select different protein-protein interaction strengths. 


Principle of Grow´n´Glow ACE1 Yeast Two-Hybrid System 


  

Vector map of Bait-Plasmid and Prey-Plasmid.


Features

  • Efficient detection of protein interactions in vivo by the copper-inducible CUP1/ACE1 system
  • Concentration can be changed to select different protein-protein interaction strengths 
  • Fewer false-positives by stringent selection for copper-resistance
  • Improved time-saving selection of positives with the well-established advantageous GFP reporter
  • No requirement for external substrates such as X-Gal when screening for positives
  • Suited for the development of high-throughput screening assays
  • Greatly reduces amount of time and effort needed to screen a cDNA library 

Applications

  • Detection of protein-protein interactions in vivo
  • Development of high-throughput screening assays
  • Gene discovery
  • Functional analysis

Contents

  • Bait plasmid, lyophilized DNA, 5 µg
  • Prey plasmid, lyophilized DNA, 5 µg
  • Bait control plasmid, lyophilized DNA, 5 µg
  • Prey control plasmid, lyophilized DNA, 5 µg
  • Positive control plasmid, lyophilized DNA, 5 µg
  • 5'-BAITprimer (0.1 nmol/µl), 500 pmol
  • 5'-PREYprimer (0.1 nmol/µl), 500 pmol
  • 3'-PREYprimer (0.1 nmol/µl), 500 pmol
  • Yeast strain ITH5, glycerol stock, 1ml

Literature