Abstract
The western blotting technique measures protein expression in a cell or tissue extract using antibody-based detection of a target protein. The antibodies used in a western blot can be specific to a total protein or to unique post-translational modifications of a protein, such as sites of phosphorylation, acetylation, methylation, or ubiquitination. The broad range of available total protein and modification-specific antibodies allows researchers to study cellular signaling events involved in many biological contexts and processes. Here we provide a comprehensive western blot protocol developed and used at Cell Signaling Technology that includes optimal reagents and support information to ensure optimal results.