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westernblot
western blot
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WESTERN BLOT - The western blot (sometimes called the protein immunoblot) is a widely used analytical technique used in molecular biology, immunogenetics and other ...
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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. * In brief, the sample undergoes protein denaturation, followed by gel electrophoresis. A synthetic or animal-derived antibody (known as the primary antibody) is created that recognises and binds to a specific target protein. The electrophoresis membrane is washed in a solution containing the primary antibody, before excess antibody is washed off. A secondary antibody is added which recognises and binds to the primary antibody. The secondary antibody is visualised through various methods such as staining, immunofluorescence, and radioactivity, allowing indirect detection of the specific target 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. |