Protein-bound polyphenols create 'ghost' band artifacts during chemiluminescence-based antigen detection
| Autor: | E.A. Foegeding, Scott M. Laster, Mary Ann Lila, Nathalie J. Plundrich |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
030103 biophysics Immunoglobulin E Horseradish peroxidase General Biochemistry Genetics and Molecular Biology law.invention Immunopharmacology & Hematologic Pharmacology 03 medical and health sciences 0302 clinical medicine Antigen law western blot artifacts General Pharmacology Toxicology and Pharmaceutics Chemiluminescence green tea polyphenols biology General Immunology and Microbiology horseradish peroxidase ghost band food and beverages Articles General Medicine protein-polyphenol interactions Primary and secondary antibodies Blot Research Note 030104 developmental biology Biochemistry Polyphenol Immunology egg white proteins biology.protein enhanced chemiluminescence Plant Biochemistry & Physiology 030217 neurology & neurosurgery Egg white |
| Zdroj: | F1000Research |
| ISSN: | 2046-1402 |
| Popis: | 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 hyperactivation of horseradish peroxidase resulting in the appearance of white “ghost” bands. 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. |
| Databáze: | OpenAIRE |
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