Detection of unusual nucleic acids with monoclonal antibodies specific for triplexes, poly(ADP-ribose), DNA-RNA hybrids, and Z-DNA.
Monoclonal antibodies which are specific for several unusual nucleic acids are now available. These include Jel 318 which is specific for triplexes, ADP-1 specific for poly(ADP-ribose), Jel 99 specific for RNA-DNA duplexes, and Jel 150 specific for Z-DNA. With the aid of these antibodies and an immunoblotting procedure, unusual nucleic acids can be detected and the amount estimated from a variety of sources. The method involves binding the nucleic acid to either nitrocellulose or Zeta Probe (a cationic nylon membrane), probing with the appropriate monoclonal antibody, followed by addition of an 125I-labeled anti-mouse second antibody. The blot is then developed by autoradiography. The technique is extremely sensitive and can be used to estimate unusual nucleic acids from crude cell extracts.
Lee
J S
JS
Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.
Latimer
L J
LJ
Sibley
J T
JT
D000911:Antibodies, Monoclonal; D001345:Autoradiography; D015152:Blotting, Northern; D015139:Blotting, Southern; D004247:DNA / Q000032:analysis*; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D009702:Nucleoside Diphosphate Sugars / Q000032:analysis*; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis*; D012313:RNA / Q000032:analysis*
D000911:Antibodies, Monoclonal; D009702:Nucleoside Diphosphate Sugars; D011064:Poly Adenosine Diphosphate Ribose; D012313:RNA; D004247:DNA