In-gel Fluorescence

In gel-fluorescence has become popular for validating the generation of fusion proteins, analyzing protein-protein interactions, and electrophoretic mobility shift assays. Fluorescent fusion proteins can easily be visualized without having to transfer to a membrane for traditional Western analysis. In the image below, fluorescent proteins are detected in a native gel. The fluorescence can then imaged with a digital imager, such as the 400, 600, or Sapphire Biomolecular Imager.

Another application of in-gel fluorescence is staining the total protein in a gel before transfer for Western blotting. The AzureRed protein stain can also provide fluorescent staining of total protein in your gel post-electrophoresis, for relative quantitative comparisons. It can be imaged with CCD- or laser-based fluorescent imaging systems and is compatible with downstream Western blotting. By including known concentration standards and imaging the stained gel, you can get quantitative information about your gel bands.