Blotto-Tween in TBS is widely used in molecular biology and biochemistry as a blocking and washing buffer in immunoassays, particularly Western blotting. It helps reduce non-specific binding of antibodies to the membrane, ensuring clearer and more accurate results. The combination of Blotto (non-fat dry milk) and Tween 20 in Tris-buffered saline (TBS) creates an effective solution for blocking unoccupied binding sites on the membrane. This minimizes background noise and enhances the specificity of antibody-antigen interactions. It is also used for diluting primary and secondary antibodies, providing a stable environment for antibody binding. Additionally, it serves as a washing buffer to remove unbound antibodies and other reagents between incubation steps, improving the overall sensitivity and reliability of the assay.