H2O2 as a DNA fragmenting agent in the alkaline elution interstrand crosslinking and DNA-protein crosslinking assays.
A method for DNA fragmentation by H2O2 in the DNA alkaline elution procedure is described. Treatment of cell suspensions for 1 h with 100 microM H2O2 or 5 mM H2O2 at 0-1 degree C resulted in DNA breakage equivalent to doses of 300 and 3000 rad of gamma-rays, respectively. The elution profiles were reproducible and H2O2 was used for measurements of interstrand crosslinks and DNA-protein crosslinks induced in HeLa cells by mitomycin C, cis-diamminedichloroplatinum(II), and trans-diamminedichloroplatinum(II). The comparison of data obtained with the use of H2O2 and gamma-rays has shown that both methods have similar sensitivity and reproducibility.
Szmigiero
L
L
Department of General Chemistry, School of Medicine in Lodz, Poland.
Studzian
K
K
D004247:DNA / Q000032:analysis / Q000187:drug effects* / Q000528:radiation effects; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000032:analysis; D005720:Gamma Rays; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide; D006863:Hydrogen-Ion Concentration; D008937:Mitomycins / Q000494:pharmacology
D004268:DNA-Binding Proteins; D008937:Mitomycins; D004247:DNA; D006861:Hydrogen Peroxide