Non-specific protein blocking is a crucial step in various molecular biology and immunoassay techniques, including Western blotting and immunohistochemistry. It involves the pre-treatment of a biological sample or a membrane to prevent non-specific binding of antibodies or other detection molecules to unwanted proteins or surfaces. This blocking step typically utilizes a blocking agent such as bovine serum albumin (BSA), non-fat milk, or serum from the same species as the secondary antibody. These blocking agents saturate unoccupied binding sites on the surface, creating a barrier that reduces the chances of non-specific interactions. By effectively “blocking” these sites, non-specific protein blocking enhances the specificity and sensitivity of assays, enabling researchers to obtain accurate and reliable results by ensuring that antibodies or detection reagents primarily bind to their intended target proteins, rather than adhering indiscriminately to other components in the sample or on the membrane.
Western blotting is a powerful, tried-and trusted technique for studying proteins for more information about experimental design see our guide