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The History and Invention of Western Blotting

Porter Birk by Porter Birk
November 6, 2021
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The History and Invention of Western Blotting
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The western blotting technique today is a staple of DNA analysis and is performed in labs worldwide. However, its origins are based, like much science, on the enhancements of previous work.

In the case of the blot, there are also some, directional puns in the naming, showing even scientists have a sense of humor.

Edwin Southern, a scientist at the University of Edinburgh, developed a new analysis to determine DNA concentration, identity and size in 1975.

The technique was called the Southern blot. It used electrophoresis to separate fragments of DNA by size and transfer them to a membrane. A radio-labeled DNA probe was used to look for specific DNA sequences within a sample.

A Stanford University group led by George Stark that includes James Alwin and David Kemp developed a similar technique in 1977. The Stark group also used a radio-labeled DNA probe, but to detect a specific RNA molecule within an RNA sample. The scientists called the method the northern blot.

Meanwhile, W. Neal Burnette was a post-doc working in the lab of Robert Nowinski at the Fred Hutchinson Cancer Research Center in Seattle. Burnett was looking to identify specific antigens within a protein mixture. He had tried unsuccessfully to combine electrophoresis with a radioimmunoassay but could not see the interactions among antibodies and separated proteins.

Burnette took inspiration from the Southern blot technique. He realized that using nitrocellulose paper, instead of chemically modified paper, facilitated the blotting of proteins better. The technique allows for clear radiographic images of antibody-specific antigens and avoided the use of a second radio-labeled antibody.

Burnette had been aware of the Southern blot and sought to honor it, and the location of his work, by naming his technique the western blot. However, when he went to publish his work in 1979, it was rejected, based on the claim that his technique was obvious, did not contribute to the scientific literature, and, surprisingly, was named in part in jest.

Despite that rejection, Burnette’s western blotting technique quickly became popular, thanks to the distribution of pre-prints to friends, who replicated the process repeatedly. The popularity of the technique spread quickly, and Burnette was soon doing daily seminars over the telephone on his blotting technique. His work was formally published in 1981 in Analytical Biochemistry, but by then was well known in the scientific community.

Controversy over Credit

The naming conventions were hardly the only controversy regarding western blotting. Over the years there has been some controversy about where credit for the technique should lie.

Harry Towbin and his colleagues at the Friedrich Miescher Institute in Switzerland had created a similar technique that derived inspiration from the northern blot. It similarly used an electrophoretic transfer of proteins from a gel onto a membrane. That team did have its work published in September 1979.

Stark and the Stanford team published in July 1979 about a similar technique that involved the passive transfer of proteins.

Burnette was aware of both labs’ work but believed his technique was simpler and universal. What he saw as strengths however, were seen by the paper’s reviewers, as weak.

Today, the naming convention persists, with scientists referring to the eastern blot, southwestern blot and the far-eastern blot, for example, as homages to the work done in the 1970s.

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