Radioimmunoassay of DNA Damaged by Ultraviolet Light
Quantitative imunoassays such as radioimmunoassay (RIA) and enzyme-linked immunosorbant assay (ELISA) are sensitive and reliable procedures used to measure UV photoproducts in purified sample DNA (Eggset et al., 1983; Strickland, 1985; Mitchell and Nairn, 1989; Matsunaga et al., 1990; Wani and Arezina, 1991; Vink et al., 1993). Competitive immunoassays have distinct advantages over other procedures in the analysis of DNA damage in human and environmental samples. For instance, unlike spectroscopic analyses, immunoassays are not limited to certain classes of adducts, and unlike HPLC and 32P-postlabeling, immunoanalysis of DNA damage does not require hydrolysis of sample DNA which can decrease the signal-to-noise ratio and alter chemical structure. ELISAs and RIAs require minimal sample manipulation and have been performed on crude cell lysates. In addition, sample DNA does not require prelabeling with a radioactive tracer and antibody binding is quasi-independent of molecular weight (i.e., DNA degradation). Immunoassays are thus readily applied to a variety of biological materials and have typically been used to measure DNA damage in cell and organ cultures, normal tissues and tumor biopsies, and various other samples including buccal cells, bone marrow aspirates, and peripheral blood lymphocytes. RIA is a sensitive and facile technique for measuring genotoxic damage in DNA.
KeywordsFluence Rate Normal Rabbit Serum Buccal Cell Antiserum Dilution Thymine Glycol
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