A FILTER BINDING ASSAY FOR CHARACTERIZATION OF ANTIBODIES TOWARDS DAMAGED DNA |
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| Authors: | Dag E. Helland Guri Eggset Hans Krokan |
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| Affiliation: | Laboratory of Biotechnology, University of Bergen, Bergen, Norway;Institute of Medical Biology, University of Tromsø, N-9001 Tromsø, Norway |
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| Abstract: | A nitrocellulose filter binding assay was used to characterize rabbit antibodies specific for UV-irradiated DNA. The dissociation constant for the UV-DNA-antibody complex was found to be 4.2 × 10−12 M. No significant binding to unirradiated DNA was observed. Unlabelled single-stranded and double-stranded DNA competed with equal efficiency for the radioactively labelled antigen (UV-irradiated φX174 DNA). The antibodies also bound to OsO4-treated DNA, suggesting that these polyclonal antibodies also recognize minor photoproducts. Caffeine efficiently decreased binding of the antibodies to UV-irradiated DNA. |
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