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Methods Mol Biol. 2012;920:163-75. doi: 10.1007/978-1-61779-998-3_12.

Immuno-slot blot assay for detection of UVR-mediated DNA damage.

Author information

1
Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester, UK. mk232@le.ac.uk

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.

PMID:
22941603
DOI:
10.1007/978-1-61779-998-3_12
[Indexed for MEDLINE]

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