Determination of the molecular weight of DNA-binding proteins using UV-crosslinking and SDS-PAGE
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Last Update: 2021-03-01
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Source: Internet
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Author: User
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We describe the use of UV-crosslinking in combination with
SDS
-PAGE to determine the approximate molecular weight of
DNA
-binding proteins. A 5-bromo-2′-deoxyuridine (5-BrdU)-substituted, radioactively labeled double-stranded oligonucleotide representing the protein binding site is incubated with a crude nuclear extract containing the protein of interest. Following irradiation with a UV light source, the DNA/protein complex is subjected to SDS-PAGE and its molecular weight determined by comparison with appropriate protein standards.
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