Western blotting is a decades-old laboratory technique that is used to detect specific proteins from cell culture, tissue, or blood specimen. The term “western blot” is a twist on the Southern blotting method developed by Edwin Southern, which is used to detect DNA and shares methodological similarities with western blotting. The western blot method was first described by Harry Towbin in 1979 but the term “western blot” was coined by W. Neal Burnette in 1981,. Since its initial description, western blotting has been used in all fields of biological and biomedical research because it is a straightforward and robust method for detecting specific proteins. Here we provide an overview of the western blot method and highlight its current applications in preclinical oncology research.