<p>(A) Total promastigote cell extracts were separated in the first dimension with a pH gradient of 3–10 (7 cm strips) followed by 12% SDS-PAGE. The separated proteins were electroblotted onto nitrocellulose membranes and probed with different pools of sera from (B) healthy dogs (n = 5; 1:500), (C, E) asymptomatic dogs (n = 5 and n = 4, respectively; 1:500), and (D, F) symptomatic dogs (n = 3 and n = 2, respectively; 1:1,000).</p