Autofluor represents the first water soluble scintillation phosphor to be developed and applied directly for use as an autoradiographic image intensifier. Autofluor rapidly penetrates acrylamide gel systems and maximizes energy transfer from labeled compound to phosphor. Autofluor contains no dimethylsulfoxide or acidic aromatic solvents. Therefore, the hazards of use related to these materials are eliminated. The band distortion that is associated with using nonaqueous enhancers is also eliminated.
In the figure to the right, the gel on the left (7%, 1mm) was dehydrated in DMSO (dimethylsulfoxide) for one hour, then impregnated in PPO-DMSO for one hour and precipitated and dried. The gel on the right was impregnated with Autofluor for one hour and dried. Both gels were exposed for 24 hours at -76°C on Kodak XR-5 X-OMat film. The single tritiated band contains 5000 dpm. Note the higher degree of resolution and band discrimination with Autofluor vs PPO-DMSO. The Autofluor procedure is the shortest and easiest procedure yet developed for enhancement and visualization of beta-emitters. In an independent test comparing eight different fluorographic methods for the detection of
35S-labeled proteins in polyacrylamide gels, Autofluor was the most effective. With Autofluor, the dpm/mm
2 required to half-saturate the x-ray film was 1/8 that required by autoradiography alone.
Storage: Autofluor has a shelf life of at least one year stored at room temperature, out of direct sunlight. Keep from freezing. At temperatures less than 20°C precipitation of water soluble phosphors may occur. Warming to approximately 30°C will redissolve the phosphors.
Protocol
Equipment 