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Quantification of photoproducts in mammalian cell DNA using radioimmunoassay.

Methods in molecular biology (Clifton, N.J.) (2006-05-06)
David L Mitchell
RESUMEN

Over the past 20 yr, the use of polyclonal and monoclonal antibodies to quantify damage in DNA has burgeoned. Immunoassays offer distinct advantages over other anaytical procedures currently used to measure DNA damage including adaptability, sensitivity and selectivity. This combination of attributes allows for the development of powerful analytical techniques to visualize and quantify specific types of DNA damage in cells and organisms exposed to subtoxic levels of xenobiotics with distinct advantages over the other procedures in the analysis of DNA damage in human and environmental samples. Radioimmunoassay (RIA) is readily applied to a variety of biological materials and has typically been used to measure DNA damage in cell and organ cultures, tissue sections and biopsies, buccal cells, bone marrow aspirates, peripheral blood lymphocytes, and urine. Here we describe the use of a very sensitive RIA for the specific quantitation of cyclobutane dimers and (6-4) photoproducts in DNA extracted from mammalian cells and tissues.

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Sigma-Aldrich
Albumin methylated from bovine serum
Sigma-Aldrich
Goat Anti-Rabbit γ-Globulin, lyophilized, Calbiochem®