Monoclonal antibody against DNA adducts with osmium structural probes.
Osmium tetroxide complexes with nitrogen ligands (Os,L) have been widely used as probes of the DNA structure. A monoclonal antibody OsBP7H8 against DNA adducts with Os,L was produced in mice. OsBP7H8 does not bind to proteins or total yeast RNA modified with Os,2,2'-bipyridine (bipy) nor to the unmodified nucleic acids and proteins. The antibody recognizes DNA modified with Os,bipy (DNA-Os,bipy) or with OsO4,1,10-phenanthroline (DNA-Os,phen) but it does not cross-react with oxidized DNA and with DNA adducts of osmium tetroxide complexes with other ligands (such as pyridine, TEMED and bathophenanthroline disulfonic acid). The affinity of OsBP7H8 to DNA-Os,phen is about five-fold higher as compared to DNA-Os,bipy. The antibody can be thus applied either for recognition of single-stranded and distorted regions in DNA (after DNA modification with Os,bipy) or for detection of both single-stranded and double-stranded DNAs (after DNA modification with Os,phen). A new simplified procedure for the dot-blot analysis is proposed, not requiring the purification of DNA-osmium adduct prior to its application to the membrane.
Bůzek
J
J
Institute of Biophysics, Academy of Sciences of the Czech Republic, Královopolská.
Kuderová
A
A
Pexa
T
T
Stanková
V
V
Lauerová
L
L
Palecek
E
E
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D002417:Cattle; D002645:Chickens; D004247:DNA / Q000276:immunology*; D018736:DNA Adducts; D006657:Histones / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009993:Osmium Tetroxide / Q000276:immunology; D011071:Poly T / Q000276:immunology; D011089:Polydeoxyribonucleotides / Q000276:immunology; D012331:RNA, Fungal / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D018736:DNA Adducts; D006657:Histones; D011089:Polydeoxyribonucleotides; D012331:RNA, Fungal; D011071:Poly T; D012710:Serum Albumin, Bovine; D004247:DNA; D009993:Osmium Tetroxide