FIG 2.

Two-dimensional profile and immunoproteomic analysis using amastigote-like total extracts of Leishmania (Viannia) braziliensis. 2DE was performed with total protein extracts (150 μg) of amastigote-like parasite forms (first dimension, IEF, pH range 4 to 7; second dimension, 12% SDS-PAGE). A stained gel is shown in panel A. Blots prepared from equivalent gels were revealed after incubation with pools of sera from mucosal (B) or cutaneous (C) leishmaniasis patients, as well as with pools of sera from noninfected subjects (D) or from patients developing Chagas disease (E). Bound antibodies were detected with conjugated goat anti-human IgG antibody at a 1:5,000 dilution. The x axis represents the tentative isoelectric point (pI), while the y axis represents the approximate molecular mass (kDa), as determined by a commercial 2DE gel marker (BenchMark protein ladder). The 39 spots identified after their specific recognition by antibodies in sera of TL patients are marked in a 2DE-stained gel shown in panel A.