A Western blot, also known as an immunoblot, is a widely used laboratory technique employed in molecular biology and immunology to detect specific proteins in a given sample. It involves three main steps: separation of proteins by size through gel electrophoresis, transfer of the proteins onto a solid support, and detection of the target protein using specific antibodies.
The Wester Western blot technique takes its name from the combining of two Western techniques: the Western blot itself and an assay called the West method. This modified protocol involves the separation of proteins using the Western blot technique, followed by specific detection of post-translational modifications, such as phosphorylation or glycosylation, using the West method.
In the West Western blot, after the proteins are separated by size using gel electrophoresis, the gel is transferred onto a membrane to preserve the protein pattern. The membrane is then incubated with specific antibodies that bind to the target protein or post-translational modification of interest. These antibodies are conjugated with enzymes or fluorescent markers that allow their visualization. By adding a substrate specific to the conjugated marker, the presence and amount of the target protein or modification can be detected and quantified.
The West Western blot technique is extensively used in research and diagnostics to study protein expression and post-translational modifications. It plays a vital role in fields such as cancer research, molecular biology, immunology, and neuroscience, providing valuable information about protein interactions, protein modifications, and protein biomarkers.
