One lab technique that has been around since what feels like the dawn of time is Western blotting, and it doesn’t look like it's going away anytime soon. Utilized by scientists worldwide, the Western blot has become an essential and powerful technique in a scientist's toolkit. One of the key players in its legacy is the Amersham Enhanced Chemiluminescence (ECL), the original ECL first introduced in the 1980s. Amersham™ ECL is still sold and can be found on many lab benches today, providing excellent and reliable results.
A quick dip into Western blotting history
In 1979, when Western blotting was first introduced, scientists found themselves spending long hours in dark rooms developing film to capture their results1. With the subsequent introduction of the Amersham ECL, researchers were able to detect small quantities of specific proteins in a sample, which was a game-changer for molecular biology. In the 1990s, early digital imaging instruments became an alternative to film, providing more reliable, reproducible, and fast results. Since then, Western blotting has been mentioned in over 400,000 PubMed-listed publications and continues to provide the foundation of protein detection and analysis in molecular biology and proteomics2.
Still blotting after all these years
With the rise of alternative methods like the enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, and mass spectrometry, many thought that Western blotting would become outdated and ultimately be replaced. Yet despite these advances, Western blotting remains resilient and continues to be a lab workhorse that keeps delivering. Scientists simply can’t part with its ease of use, and the fast, cost-effective, and reliable results it provides, crucial for validating antibody specificity, detecting post-translational modifications, and confirming proteomics data3.
Lights, cameras, detection: The imaging evolution
One aspect of Western blotting that continues to evolve is the techniques used for imaging. While X-ray film was once the standard, today researchers more commonly rely on digital imaging solutions, including charge-coupled device (CCD) camera-based imagers and scanner-based systems. Modern tools like Amersham™ ImageQuant™ 800 Western blot imaging systems from Cytiva are CCD-based imagers, featuring sensitive Fujifilm optics, and the signal-to-noise optimization watch (SNOW) imaging mode, which automatically provides sharp images with the best signal-to-noise ratio. These systems enhance sensitivity by detecting weaker bands in Western blots while also improving efficiency, automatically capturing the best possible image, freeing up researchers' time by eliminating the need for optimizing exposure and capture settings.
Consistent Western blots every time
While Western blotting remains a lab favorite, scientists know the challenge of reproducibility4. One day, the bands are crisp and sharp, and the next day, they could appear as a merged smear. Modern imaging systems like the Amersham™ ImageQuant™ 800 and Amersham™ Typhoon™ laser scanner from Cytiva help researchers tackle these challenges, with their precise, standardized imaging protocols, and robust analysis tools. The Amersham ImageQuant 800 offers high-sensitivity optics and consistent exposure settings for clear chemiluminescent and fluorescent blots, and the Amersham Typhoon scanner delivers high-resolution, multi-channel fluorescence scanning with adjustable sensitivity. Both systems feature user-friendly software for automated quantification, ensuring results are clear, reliable, and comparable across experiments. By minimizing variability, modern imaging platforms support more reproducible and trustworthy results.
Conclusion: Old dog, new tricks
Western blotting has certainly earned its place in the hall of fame, not just for the results it provides, but also for its adaptability. With tools like Amersham ECL still shining and new advanced imaging technology making imaging sharper and smarter than ever, Western blotting isn’t going anywhere anytime soon.
Check out guide for Western Blotting from Cytiva success and explore the full range of available instruments and consumables to discover how you can achieve clearer, more reproducible, and consistent blot images.
Sources
1. Sule R, Rivera G, Gomes AV. Western Blotting (immunoblotting): History, Theory, Uses, Protocol and Problems. BioTechniques. 2023;75(3):99-114. doi:10.2144/btn-2022-0034
2. Moritz CP. 40 years Western blotting: A scientific birthday toast. J Proteomics. 2020;212:103575. doi:10.1016/j.jprot.2019.103575
3. Yang PC, Mahmood T. Western blot: Technique, theory, and trouble shooting. North Am J Med Sci. 2012;4(9):429. doi:10.4103/1947-2714.100998
4. Kroon C, Breuer L, Jones L, et al. Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them. Bertrand MJM, ed. PLOS Biol. 2022;20(9):e3001783. doi:10.1371/journal.pbio.3001783