Abstract
Western blotting is a well establish technique used for the routine detection of specific proteins from complex mixtures. The immunodetection step of western blotting is a long and tedious process, involving multiple changes of solutions, and is usually extended over several hours to overnight. This paper describes an innovative, vacuum‐based immunodetection approach, which allows the detection of the protein in less than 45 minutes. The system can be used for rapid antibody optimization studies, antibody screening, simultaneous detection of different proteins in multiples blots, or routine western blot analysis. Vacuum filtration enables efficient blocking and washing of the membrane, speeds up the overall process and make the system easy to use. Blocking of the membrane occurs rapidly in the presence of low concentration of non fat dry milk or other common blocking reagents. Since incubation steps are typically performed in ten minutes, users can sequentially process several blots in a day. The system in compatible with PVDF or nitrocellulose membranes and detection can be accomplished by any chemiluminescence or fluorescence detection method. The SNAP id™ system is a novel immunodetection system that uses the basic steps of the standard immunodetection protocol, is easy to use and offers a substantial reduction in processing time.This research has been totally funded by Millipore Corporation.
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