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

Full Title: Protein-bound polyphenols create “ghost” band artifacts during chemiluminescence-based antigen detection

Journal: F1000Research

Year of Publication: 2017

PHHI Author(s): Mary Ann Lila
Publication Author(s): Nathalie Plundrich, Mary Ann Lila, Edward Foegeding, Scott Laster

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

Link to Article: https://www.ncbi.nlm.nih.gov/pubmed/28721200