Western blotting is a technique used routinely by many laboratories for protein identification and analysis. The procedure involves many steps that must be performed with great accuracy and efficiency. Briefly, a typical protocol begins with an experimental procedure that ends up with a sample containing protein molecules, which must be characterized in order to reach to a conclusion. A simple way to accomplish this analysis is to separate the protein mixture by electrophoresis (SDS-PAGE for example) and to transfer these proteins to an appropriate scaffold (such as nitrocellulose), on which the final detection step will take place. Due to the many steps that must be accomplished in order to reach to the final image, Western blot analysis can be considered to be a form of art. The final step of the protocol involves the detection of the antibody bearing an enzyme (such as horseradish peroxidase) that modifies an appropriate substrate so that it emits light. Traditionally, the emitted light was detected using X-ray film and a series of solutions in a darkroom. This detection method cannot be considered to be ideal because it introduces a few more steps at which a possible mistake can ruin the entire experiment. Furthermore, the traditional film can introduce more complexity as it requires careful handling in the darkroom and the possession of more equipment to scan the image and transform it into digital form.
A brilliant alternative to the above detection method is the Luminescent Image Analyzer LAS-3000 offered by Fujifilm Life Science. This instrument has a CCD camera with astonishing sensitivity. Furthermore, the LAS-3000 can be used for a wide variety of applications such as fluorescence, chemifluorescence, chemiluminescence and digitization. The resolution that can be achieved using LAS-3000 peaks at 6.3 Megapixels. The device is accompanied by software that is extremely easy to learn and to use routinely.
After incubating your membranes with the appropriate antibodies you can place them centered on the tray that accompanies LAS-3000 and then detection can be complete in a matter of minutes without any further complicated steps. Forget the darkroom, and the drying of the film. Your image is ready in a matter of minutes in a digital form that can be stored in a portable device right away.
The sensitivity of the lenses can give you astonishing detail of detection especially when using substrates such as SuperSignal West Pico Chemiluminescent Substrate (from Pierce), which further enhance the detection power of Western blots.
Overall, I am extremely satisfied with the performance of this device and honestly, I haven’t been able to find any major flaws with it. I recommend it to anyone who wants to detect extremely low amounts of proteins without having to worry about the performance of their detection systems at the final step of the process, after a long day in the laboratory.
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Sardis Marios Frantzeskos, BS
Graduate Student
Department of Biology
University of Crete
Greece
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