I have done a western blot and I want to remove the background signal when doing densitometry analysis of my protein bands of interest. I have read some articles online such as this one regarding high background in western blots. I have learnt that there are many factors that can contribute to high background in a western blot. However, I am still not sure what exactly is defined as background. Does it refer to non-specific antibody binding on the membrane, or can it also refer to regions on the membrane where there is no binding to any protein?
Any insights are appreciated.