Calibration of target amounts of DNA in hybridization experiments using monoclonal anti-nucleoside antibodies.
Two anti-nucleoside monoclonal antibodies (A-16 and G-K21) were raised after immunizing mice with adenosine or guanosine coupled to bovine serum albumin by periodate oxidation. They were selected for their ability to detect these immunogens and single-stranded DNA in an enzyme-linked immunosorbent assay test. The antibodies were purified from ascitic fluids, their isotypes were determined and their ability to detect DNAs and RNAs on nitrocellulose membranes was tested. They belonged to the IgG1 subclass and were both able to recognize picogram amounts of single-stranded DNAs on nitrocellulose sheets, whatever the origin of the nucleic acid, but were unable to detect RNA efficiently. The same monoclonal antibodies were used to estimate minute amounts of target staphylococci DNAs to permit standardization of non-radioactive hybridization experiments for detection of antibiotic resistance genes.
Traincard
F
F
Laboratoire des Sondes Froides, Paris, France.
Sakamoto
H
H
Rouyre
S
S
Mazie
J C
JC
Guesdon
J L
JL
D000241:Adenosine / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D004247:DNA / Q000032:analysis* / Q000276:immunology; D004277:DNA, Single-Stranded / Q000276:immunology; D006151:Guanosine / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D009693:Nucleic Acid Hybridization
D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded; D007074:Immunoglobulin G; D006151:Guanosine; D004247:DNA; D000241:Adenosine