Application Note: Identification of Co-migrating Proteins on the FluorChem M System

Western blotting is an essential tool in protein research. Traditionally, proteins are labeled with secondary antibodies tagged with chemiluminescent substrates and are detected using photographic film. Chemiluminescent Western blotting has been a dominant method of detection for identification of proteins in cell or tissue samples for decades. While it offers significant advantages over other methods it also has certain limitations, one being the ability to analyze multiple proteins on the same blot. Fluorescent imaging technology gives researchers the ability to move beyond the traditional Western blot assay by offering the flexibility to multiplex and detect co-migrating proteins. Native and phosphorylated isoforms can be probed simultaneously and quantified without the additional time and signal loss in sequential stripping and re-probing.