Is Phosphorylated Protein Detection More Efficient by Fluorescent Western Blot?

The detection of phosphorylated proteins usually requires showing both nonphosphorylated and phosphorylated species. For traditional chemiluminescent methods, the negligible size difference between the phosphorylated and nonphosphorylated protein of interest requires probing with a phospho-specific antibody, then stripping, and then re-probing with the protein-specific antibody. Along with potential negative effects on data quality, the process of stripping and re-probing can take a considerable amount of time and reagents. This application note demonstrates the comparison of a chemiluminescent detection protocol and a protocol using fluorescent antibodies.