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immunoblottings
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| A technique for analysing or identifying proteins in a mixture, involving separation by electrophoresis followed by staining with antibodies. |
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The Western blot (sometimes called the protein immunoblot) is a widely used analytical technique used in molecular biology, immunogenetics and other molecular biology disciplines to detect specific proteins in a sample of tissue homogenate or extract. * Synthetic or animal-derived antibodies are created that react with a specific target protein. The sample material undergoes protein denaturation, followed by gel electrophoresis. Next, the electrophoresis membrane is washed in a solution containing the specific antibody. The excess antibody is then washed off, and a secondary antibody that reacts with the first antibody is added. Through various methods such as staining, immunofluorescence, and radioactivity, the secondary antibody can then allow visualisation of the protein. * Other related techniques include dot blot analysis, quantitative dot blot, immunohistochemistry, and immunocytochemistry where antibodies are used to detect proteins in tissues and cells by immunostaining, and enzyme-linked immunosorbent assay (ELISA). * The name western blot is a play on the eponymously-named Southern blot, a technique for DNA detection developed earlier by Edwin Southern. Similarly, detection of RNA is termed northern blot and was developed by James Alwine, David Kemp, and George Stark at Stanford. The term "western blot" was given to the technique by W. Neal Burnette, although the method itself originated in the laboratory of Harry Towbin at the Friedrich Miescher Institute. |