Immunoblot analysis of the reaction of sera from vaccinated patients against PSMA. LNCaP lysates (A) or recombinant PSMA (B) were run on 12% polyacrylamide gel, transferred onto nitrocellulose, and treated with serum from a healthy male donor or from immunized patients who had or had not developed antibodies against the target antigen. (A) Lane 1, LNCaP lysate pretreated with serum from a healthy male donor diluted 1:50; lane 2, LNCaP lysate pretreated with a negative patient’s serum diluted 1:50; lane 3, LNCaP lysates pretreated with a positive patient’s serum diluted 1:50. (B) Lane 1, recombinant PSMA pretreated with serum from a healthy male donor diluted 1:50; lane 2, recombinant PSMA pretreated with a negative patient’s serum diluted 1:50; lane 3, recombinant PSMA pretreated with a positive patient’s serum diluted 1:50. After pretreatment all blots were treated consecutively with specific rabbit anti-PSMA antibodies and antirabbit IgG serum labeled with AP.