How it works

 

PROTEIN INTERACTION ANALYSIS SERVICE

experimentally validated
full customization
cost-efficient
comprehensive reporting

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The PROCOMCURE Biotech Y2H protein/peptide interaction analysis is based on the reconstitution of a functional transcription factor (TF) followed by the expression of a reporter gene in genetically modified, auxotrophic yeast cells. Upon physical binding of the protein of interest (bait) to a protein fragment or full-length protein (prey), the DNA-Binding Domain (BD) of the TF is brought in close proximity to its Activation Domain (AD). Reconstitution of the functional TF activates the transcription of reporter genes, which allows yeast cells to grow on a selective medium. Auto-activators and false-positive clones are eliminated in the quality control process. The plasmid DNA of positive clones is then sequenced and analyzed to identify the interaction partners.

 

  • Setup: Choose between the various full-length or fragment libraries and submit your protein of interest (protein sequence or protein-ID is sufficient)
  • Construct design: The DNA sequence of your target protein will be designed and cloned into Y2H vectors ready for screening
  • Interaction screening: An perfected (automated, high-throughput configuration) protein interaction screen will be performed to identify interaction partners for your protein of interest (first hits)
  • Confirmation: Extended reanalysis of hits (candidate proteins) resulting from the interaction screen (reproducibility, false-positives elimination) and subsequent DNA-sequencing validate the reliability of your hits
  • Validation (optional): Immune precipitation, pull-down analysis, biolayer interferometry – surface plasmon resonance (SPR), FRET, fluorescence polarization, alpha screening, co-localization available (please contact)
  • Results: Documents contain a brief summary as well as a full documentation of the PROTEIN INTERACTION ANALYSIS SERVICE.