Photoimmunodetection: a nonradioactive labeling and detection method for DNA.
We describe a simple detection system for DNA based on antibody detection of UV-induced photoproducts. It includes a convenient and inexpensive labeling procedure, which is completed in 5-10 min. The only equipment required is a UV source such as an ordinary transilluminator or a DNA crosslinker. Using a monoclonal antibody specific for thymine dimers, coupled to horseradish peroxidase, we are able to detect subpicogram amounts of UV-irradiated DNA directly, and approximately 10 pg homologues DNA indirectly by hybridization with an irradiated probe.
Marvik
O J
OJ
Biotechnology Centre, University of Oslo, Norway.
Isaksen
M L
ML
Roza
L
L
Lindqvist
B H
BH
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D010584:Bacteriophage phi X 174 / Q000235:genetics; D003432:Cross-Linking Reagents; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D004277:DNA, Single-Stranded / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics; D007124:Immunoenzyme Techniques; D009693:Nucleic Acid Hybridization; D011740:Pyrimidine Dimers / Q000032:analysis / Q000276:immunology; D014466:Ultraviolet Rays
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D015342:DNA Probes; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D011740:Pyrimidine Dimers; D004247:DNA