Analysis by ELISA and western blotting of trypanosome antigen specific antibodies in sera from intact and <i>Prf1</i>^<i>-/-</i> C57BL/6 mice infected for up to 30 days with <i>T</i>. <i>brucei</i> AnTat 1.1.

<p><i>T</i>. <i>b</i>. <i>brucei</i> AnTat 1.1 lysate (1 μg protein in 0.05% Np40/well) was coated on wells of an ELISA plate and used to detect trypanosome antigen specific antibodies of: (<b>A</b>) IgM, <b>(B)</b> IgG, <b>(C)</b> IgG1, <b>(D)</b> IgG2a classes in dilutions of serum (doubling dilutions from 1:1000 to 1:54,000) from groups of uninfected (naïve) intact C57BL/6 (gray dots) and <i>Prf1</i>^<i>-/-</i> C57BL/6 mice (black squares) and mice that had been infected for 10, 20 or 30 days with <i>T</i>. <i>brucei</i> AnTat 1.1(n = 6/group). Trypanosome lysate (0.5% NP40) was subjected to SDS-PAGE, transferred to membrane and stained with pooled serum from a group of uninfected intact or <i>Prf1</i>^<i>-/-</i> mice (n = 6), or mice infected with <i>T</i>. <i>brucei</i> AnTat 1.1 for 10, 20 or 30 days as indicated (n = 6/group) to detect polypeptide-specific antibodies of: <b>(E)</b> IgM, <b>(F)</b> IgG1 and <b>(G)</b> IgG2a classes. Results are representative of results obtained in individual mice of each group in 3 identical experiments.