A method is described for selecting monoclonal antibodies (mAb) agains
t DNA-binding protein, The protocol involves a non-radioactive solid-p
hase 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 wi
th crude cell extract as the antigen, MAbs obtained by this method cou
ld supershift the DNA-protein complex in the electromobility shift ass
ay, and were sufficient for immunoscreening of a cDNA expression libra
ry.