DNA adducts in mammalian cells exposed to N-acetoxy-2-acetylaminofluorene (NA-AAF), an activated derivative of the potent carcinogen 2-AAF, play significant roles in cell killing, chromosome aberration, gene mutation and neoplastic transformation. NA-AAF binds covalently to guanine in the DNA of mammalian cells and produces three different DNA adducts. The C-8 adducts dG-C8-AAF and deacetylated dG-C8-AF account for the major portion of the DNA-bound products, while the minor N2 adduct dG-N2-AAF accounts for the remainder. The relative induction levels of the two major C-8 adducts vary among cell types. These adducts distort the DNA helix and therefore are repaired by nucleotide excision repair in human cells. Our AAF-1 antibodies bind most efficiently to dG-C8-AAF and less efficiently to dG-C8-AF in denatured DNA. The antibodies enable one to detect AAF-DNA adducts in DNA from cultured cells using an enzyme-linked immunosorbent assay (ELISA) and to visualize them in cultured cells or rodent tissues by immunofluorescence (IF). This technology will contribute to understanding of molecular mechanisms in AAF-related research fields including cancer research, anticancer research and toxicology.

Source: Toshio Mori Professor, Research Institute for Advanced Medicine, Nara Medical University.

References:
1) R.H. Heflich and R.E. Neft, Genetic toxicity of 2-acetylaminofluorene, 2-aminofluorene and some of their metabolites and model metabolites. Mutation Res. 318 (1994) 73-174.
2) E.Kriek, Fifty years of research on N-acetyl-2-aminofluorene, one of the most versatile compounds in experimental cancer research. J. Cancer Res. Clin. Oncol. 118 (1992) 481-489.
3) T. Iwamoto et al., In situ detection of acetylaminofluorene-DNA adducts in human cells using monoclonal antibodies. DNA Repair 3 (2004) 1475-1482.

DNA adducts in mammalian cells exposed to N-acetoxy-2-acetylaminofluorene (NA-AAF), an activated derivative of the potent carcinogen 2-AAF, play significant roles in cell killing, chromosome aberration, gene mutation and neoplastic transformation. NA-AAF binds covalently to guanine in the DNA of mammalian cells and produces three different DNA adducts. The C-8 adducts dG-C8-AAF and deacetylated dG-C8-AF account for the major portion of the DNA-bound products, while the minor N2 adduct dG-N2-AAF accounts for the remainder. The relative induction levels of the two major C-8 adducts vary among cell types. These adducts distort the DNA helix and therefore are repaired by nucleotide excision repair in human cells. Our AAF-1 antibodies bind most efficiently to dG-C8-AAF and less efficiently to dG-C8-AF in denatured DNA. The antibodies enable one to detect AAF-DNA adducts in DNA from cultured cells using an enzyme-linked immunosorbent assay (ELISA) and to visualize them in cultured cells or rodent tissues by immunofluorescence (IF). This technology will contribute to understanding of molecular mechanisms in AAF-related research fields including cancer research, anticancer research and toxicology.

Source: Toshio Mori Professor, Research Institute for Advanced Medicine, Nara Medical University.

References:
1) R.H. Heflich and R.E. Neft, Genetic toxicity of 2-acetylaminofluorene, 2-aminofluorene and some of their metabolites and model metabolites. Mutation Res. 318 (1994) 73-174.
2) E.Kriek, Fifty years of research on N-acetyl-2-aminofluorene, one of the most versatile compounds in experimental cancer research. J. Cancer Res. Clin. Oncol. 118 (1992) 481-489.
3) T. Iwamoto et al., In situ detection of acetylaminofluorene-DNA adducts in human cells using monoclonal antibodies. DNA Repair 3 (2004) 1475-1482.