Monoclonal anti-DNA antibody: preparation and availability.
Twelve hybridomas producing anti-DNA antibody were established using single-stranded DNA of phi x174 and alkali-denatured DNA fragments of G4 phage as an antigen. To remove high DNase activity in ascites, antibodies were purified on heparin-Sepharose column following Sephacryl S-200 column chromatography. With purified antibodies, binding specificity was determined by competitive experiment and foot-printing. Antibodies specific to different higher-order structures of DNA, such as deoxypyrimidine-cluster, stem-region in stem-loop structure and superhelical structure, were obtained. These antibodies were useful to pursue conformational changes of DNA in aqueous solution and in interaction with protein during transcription and replication. Topography of DNA in DNA-virus infected as well as uninfected cells was also defined with these monoclonal anti-DNA antibodies by indirect immuno-fluorescence method.
Fukada
M
M
Department of Microbiology, Aichi Medical University, Aichi, Japan.
Yoshikawa
K
K
D000818:Animals; D000974:Antibodies, Antinuclear / Q000032:analysis* / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D010584:Bacteriophage phi X 174 / Q000235:genetics; D002460:Cell Line; D004277:DNA, Single-Stranded / Q000032:analysis* / Q000276:immunology; D004279:DNA, Viral / Q000032:analysis* / Q000276:immunology; D005455:Fluorescent Antibody Technique; D009691:Nucleic Acid Denaturation; D013539:Simian virus 40 / Q000235:genetics
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded; D004279:DNA, Viral