Hey all,
This is less of a question and more of a hint because I've been struggling with this for almost a year during my graduate career. In case you work with proteins and phenolic compounds you may run into some issues with Western blotting/chemiluminescence detection methods. Check out our paper here from 2017 if you have noticed similar problems. May save a lot of time, money, resources and frustration!
Article Protein-bound polyphenols create “ghost” band artifacts duri...
As I understand it, proteins are first separated by electrophoresis, then specific proteins are determined by their interaction with specific antibodies. The essence of electrophoresis is the movement of electron-charged proteins under the action of an electric current
Beautiful question and great answers
I wish you success in the service of humanity and science
Rimma Mukhamatkhanova Yes, but what I am saying is, that phenolic compounds can covalently bind to proteins and survive the preparation for SDS-PAGE (heat, reducing agent etc.). This means they make it into the gel and subsequently onto the membrane you're using for subsequent detection. With the specific detection method I used using specific antibodies, I ran into detection/interference issues. Had to switch to a non-enzymatic detection method using fluorescence.
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