title: Immuno-slot blot assay for detection of UVR-mediated DNA damage.
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.
authors.affiliation: Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester, UK. mk232@le.ac.uk
authors.forename: Mahsa
authors.initials: M
authors.lastname: Karbaschi
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Brady
authors.forename: Mark D
authors.initials: MD
authors.lastname: Evans
authors.forename: Marcus S
authors.initials: MS
authors.lastname: Cooke
mesh_terms: D000818:Animals; D002138:Calibration; D002417:Cattle; D004247:DNA / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004249:DNA Damage; D003849:Deoxyguanosine / Q000378:metabolism; D015151:Immunoblotting / Q000379:methods*; D008315:Malondialdehyde / Q000378:metabolism; D014466:Ultraviolet Rays / Q000009:adverse effects*
chemical_list: D008315:Malondialdehyde; D004247:DNA; D003849:Deoxyguanosine