Abstract
<p>A complete Western blot protocol for detecting recombinant fusion proteins using epitope tag-specific antibodies (6×His, Flag, mCherry, HA, Myc, GFP). Covers sample preparation, SDS-PAGE, transfer, blocking, antibody incubation, and chemiluminescent detection. Protocol developed by abinScience / AtaGenix Technical Team. Epitope tags are short peptide sequences or fluorescent proteins fused to recombinant proteins to enable detection, purification, and tracking without requiring a target-specific antibody. This protocol provides optimized conditions for Western blot detection of the most commonly used tags — 6×His, Flag (DYKDDDDK), HA, Myc, mCherry, and GFP — in mammalian cell lysates, bacterial lysates, and purified protein samples. It includes tag-specific tips for antibody selection, blocking buffer compatibility, and troubleshooting.</p>