Quantitative measurement of ultraviolet-induced damage in cellular DNA by an enzyme immunodot assay.
A simple enzyme immunoassay procedure was developed for the quantitative determination of 254-nm uv-induced DNA damage in cells. With the use of specific antibodies to uv-irradiated DNA and horseradish peroxidase-conjugated antibody to rabbit IgG, the extent of damaged DNA in uv-irradiated rat spleen mononuclear cells was quantitatively measurable. Through the use of this method, the amount of damaged DNA present in 2 X 10(5) cells irradiated at a dose of 75 J/m2 was estimated to be 7 ng equivalents of the standard uv-irradiated DNA. In addition, when the cells, irradiated at 750 J/m2, were incubated for 1 h, the antigenic activity of DNA decreased by 40%, suggesting that a repair of the damaged sites in DNA had proceeded to some extent in the cells.
Wakizaka
A
A
Department of Biochemistry, Akita University School of Medicine, Japan.
Nishizawa
Y
Y
Aiba
N
N
Okuhara
E
E
Takahashi
S
S
D000818:Animals; D004249:DNA Damage; D003720:Densitometry; D007124:Immunoenzyme Techniques; D051381:Rats; D011919:Rats, Inbred Strains; D013154:Spleen / Q000166:cytology / Q000528:radiation effects; D014466:Ultraviolet Rays / Q000009:adverse effects*