Abstract
Solar ultraviolet radiation (UVR), through the formation of DNA photolesions, is the primary cause of most skin cancers. A better understanding of the mechanisms of UVR-induced DNA damage may help prevent skin cancer and this may be achieved using methods to quantify DNA damage. The immuno-slot blot (ISB) method is routinely used for detection and quantification of any heat- and alkali-stable DNA adducts for which a sufficiently specific monoclonal antibody is available. The main steps in ISB are fragmentation and denaturation of the DNA, immobilization of DNA to a nitrocellulose filter, incubation with primary antibody against a specific DNA adduct, incubation with an enzyme-linked secondary antibody and finally chemiluminescence detection and quantification of the DNA adducts.
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Acknowledgment
The authors would like to acknowledge Prof. Peter B. Farmer (University of Leicester, UK) for use of immuno-slot blot equipment and laboratory space, Dr. Rajinder Singh (University of Leicester, UK) for advice, and Prof. Lawrence Marnett (Vanderbilt University, USA) for supplying the M1dG antibody.
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Karbaschi, M., Brady, N.J., Evans, M.D., Cooke, M.S. (2012). Immuno-Slot Blot Assay for Detection of UVR-Mediated DNA Damage. In: Bjergbæk, L. (eds) DNA Repair Protocols. Methods in Molecular Biology, vol 920. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-998-3_12
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DOI: https://doi.org/10.1007/978-1-61779-998-3_12
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