Determination of the molecular weight of DNA-binding proteins using UV-crosslinking and SDS-PAGE |
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| Authors: | Siegmund S. Wolf Karim Roder Michael Schweizer |
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| Affiliation: | (1) Institute of Food Research, Genetics and Microbiology Department, Norwich Research Park, Colney, NR4 7UA Norwich, UK |
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| Abstract: | 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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| Keywords: | UV-crosslinking SDS-PAGE nuclear protein extract DNA-binding protein 5-bromo-2′ -deoxyuridine |
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