A solid-phase assay to screen monoclonal antibodies against DNA-binding protein.
A method is described for selecting monoclonal antibodies (mAb) against DNA-binding protein. The protocol involves a non-radioactive solid-phase DNA binding assay using a 96-well plate. Because the solid-phase assay is highly specific and sensitive, partially purified antigen is sufficient for the immunization, and mAb screening can be performed with crude cell extract as the antigen. MAbs obtained by this method could supershift the DNA-protein complex in the electromobility shift assay, and were sufficient for immunoscreening of a cDNA expression library.
Ishii
N
N
Department of Mycology, Nippon Roche Research Center (NRRC), Kajiwara 200, Kamakura 247, Japan. nobuya.ishii@roche.com
Nakayama
H
H
Katayama
J
J
Arisawa
M
M
Aoki
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001667:Binding, Competitive; D001710:Biotin; D002176:Candida albicans / Q000737:chemistry; D015342:DNA Probes; D004268:DNA-Binding Proteins / Q000276:immunology*; D004586:Electrophoresis; D006825:Hybridomas / Q000276:immunology; D007124:Immunoenzyme Techniques; D007163:Immunosorbent Techniques; D051379:Mice; D012441:Saccharomyces cerevisiae / Q000737:chemistry
D000911:Antibodies, Monoclonal; D015342:DNA Probes; D004268:DNA-Binding Proteins; D001710:Biotin