Protein-bound polyphenols create "ghost" band artifacts during chemiluminescence-based antigen detection

Mary Ann Lila, Nathalie Plundrich, Edward Foegeding, Scott Laster

Research output: Contribution to journalComment/debate

Abstract

Antigen detection during Western blotting commonly utilizes a horseradish peroxidase-coupled secondary antibody and enhanced chemiluminescent substrate. We utilized this technique to examine the impact of green tea-derived polyphenols on the binding of egg white protein-specific IgE antibodies from allergic human plasma to their cognate antigens. Our experiments unexpectedly showed that green tea-derived polyphenols, when stably complexed with egg white proteins, caused "ghost" band formation in the presence of horseradish peroxide. This study suggests that caution should be taken when evaluating polyphenol-bound proteins by enhanced chemiluminescence Western blotting using horseradish peroxidase and demonstrates that protein-bound polyphenols can be a source of "ghost" band artifacts on Western blots.

Original languageEnglish (US)
Article number254
JournalF1000Research
Volume6
DOIs
StatePublished - Jan 1 2017

Keywords

  • Egg white proteins
  • Enhanced chemiluminescence
  • Ghost band
  • Green tea polyphenols
  • Horseradish peroxidase
  • Protein-polyphenol interactions
  • Western blot artifacts

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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