FAQ: What are the advantages of the IMPACT System?

  • Single column purification - no additional steps to remove the affinity tag
  • Yields native amino acid sequence
  • Fusion of a target protein to either the C-terminus or N-terminus of the intein tag
  • Proteases are NOT required to remove the affinity tag from the target protein
  • Use of either thiol reagent or pH and temperature shift to induce on-column cleavage
  • Isolation of proteins with or without an N-terminal methionine residue
  • Production of proteins possessing an N-terminal cysteine and/or C-terminal thioester for use in protein labeling, ligation and cyclization [See Part 6: Applications: Protein ligation and labeling]
  • T7 Promoter for high-level expression and tight regulation of transcription
  • Protein semisynthesis - The ability to create a thioester at the C-terminus for ligation with a N-terminal containing cysteine peptide or tag [See Part 6: Applications: Protein ligation and labeling]
  • The ability to label the C-terminus of the target protein [See Part 6: Applications: Protein ligation and labeling]
  • IMPACT vectors pTYB21 (NEB #N6709) and pTYB22 (NEB #N6710) contain a MCS that is compatible with other NEB expression vectors.