| Southwestern Blot |
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| CATEGORIES ABOUT SOUTHWESTERN BLOT | |
| molecular biology | |
| laboratory techniques | |
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However, since the first southwestern blottings, many more have been proposed and discovered. Large amounts of Protein s and their Degradation when being isolated hampered previous protocols. "South Western blot mapping" is performed for rapid characterization of both DNA binding proteins and their specific sites on genomic DNA. Proteins are separated on a Sodium Dodecyl Sulfate (SDS), polyacrylamide gel (PAGE), renatured by removing SDS in the presence of urea, and blotted onto nitrocellulose by diffusion. The genomic DNA region of interest is digested by restriction enzymes selected to produce fragments of appropriate but different sizes, which are subsequently end-labeled and allowed to bind to the separated proteins. The specifically bound DNA is eluted from each individual protein-DNA complex and analyzed by acrylamide gel electrophoresis. Evidence that tissue-specific DNA binding proteins may be detected by this technique is presented. Moreover, their sequence-specific binding allows the purification of the corresponding selectively bound DNA fragments and may improve protein-mediated cloning of DNA regulatory sequences. EXTERNAL LINK |
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