Western blot (WB), also known as immunoblotting, works based on the following core principle: proteins are separated via SDS-PAGE electrophoresis and transferred onto PVDF or NC membranes. Leveraging specific antigen-antibody binding, the primary antibody targets the protein of interest, while HRP-conjugated secondary antibodies bind to the primary antibody. Luminescence is generated upon catalysis of ECL substrate. The molecular weight of proteins is determined by band position, and grayscale intensity represents protein abundance.