Has the Western Blot 'Evolved'?

Feb-02-2016 0 comments Cube Biosystems

There are those who believe that the history of the Western Blot is very much the present of the Western blot; that the technique hasn't especially evolved since the late 1970s. Robert T. Kennedy of the University of Michigan, for example, claims that 'the fundamentals of this technique have been around for more than 30 years and have hardly changed'. Of course, we can see where he's coming from - the technique as a whole hasn't changed significantly - but there has been noticeable evolution.

Regardless of which side of the great 'who invented it' debate you stand on, something that we can all agree on is that the Western Blot technique for protein identification and quantification has definitely stood the test of time. We've been using it as one of the primary methods for singling out specific proteins since 1979, and the Western Blot has undoubtedly had a significant impact upon many aspects of modern medicine, particularly immunology, neuroscience, and in recent drug development.

It Broke, We Fixed It

FWhat sort of images do you typically get from your Western Blots — Look all too familiar?

Back in 1979, the Western Blot was far from perfect. It's still not perfect today, but scientists have been working to identify weak areas within the technique in order to create and introduce 'fixes' which enable us to slightly adapt it to improve success. As we know, reliability and consistency are common issues surrounding the Western Blot technique. It's reported that the method fails approximately 25 percent of the time, while variability between users can average 35 to 45 percent.

So while the Western Blot, at its core, has largely remained the same, what we have seen in the last 30 years are changes being implemented that give us the option to adjust the technique - changes that mean we can identify and isolate proteins quicker, with less manual intervention, and with greater throughput, all while minimizing variability and boosting accuracy. We're well on the way, but we're still not there yet, and many more changes are expected to be introduced to improve the way we work.

The Future of the Western Blot

As the Western Blot has already proved to be the preferred method for more than 30 years, there are many who believe that we'll simply see more changes to the technique, rather than a full overhaul; that we'll keep adapting while retaining the core, rather than bring in a whole new technique. We can already see evidence of this, with the rising popularity of CCD imaging and the decline in traditional film use. We're continually making changes that enable us to use this old technique in a modern setting.

However, the Western Blot was born from both the Southern and Northern Blot techniques, so could this in turn eventually be replaced? Some are arguing 'yes', and that it's about time we had a change. In his paper 'Can Proteomics Retire the Western Blot?', Matthias Mann argues that 'biologists are currently stuck in a time warp - they basically use the same tools for protein detection as they have for the past 20 years. So what's the answer? What could we see being pushed forward as the new Western Blot?

There are many who are very passionate about mass spectrometry as an alternative to Western Blotting, and it's been referred to as a 'next-generation platform that overcomes many of the limitations of Western Blotting'. The advantage, of course, is that inaccuracies relating to antibodies are prevented, but mass spec has its flaws, too. While detection with antibodies can be achieved through a few hundred molecules, for example, many more are required for successful identification through MS.

While it's important for us to be open to alternatives, and open to progression, it appears that there are a few scientists who are striving for a change merely for the sake of it. Western Blotting isn't perfect, we know this, but from the evolution of the technique we've seen so far, we can also conclude that we can successfully streamline and modernize it, to make it as near to perfect as we can get. The Western Blot technique may be a bit old, but we don't think it's had its day just yet.

For those still stuck in the time warp, though, At VitaScientfic we offer several products from the unique Intelli Mixer, antibodies, blue-film or imagers, and, filter sandwiches that will help you achieve that publication quality western.