Immune Response Induced by an Immunodominant 60 kDa Glycoprotein of the Cell Wall of Sporothrix schenckii in Two Mice Strains with Experimental Sporotrichosis

Figure 3

Purification of gp60. Cell wall proteins of yeast-like cells of S. schenckii were separated by sequential steps of liquid phase isoelectric focusing and electrophoresis with continuous electroelution as described in Materials and Methods. After each step, fractions were analyzed by SDS-PAGE ((a), (c), and (e)) and Western blot ((b), (d), and (f)) using polyclonal anti-gp60 primary antibodies and peroxidase-conjugated secondary antibodies. Signal was revealed with a DAB-peroxidase substrate solution. Fractions separated after the first ((a) and (b)) and second ((c) and (d)) liquid phase isoelectric focusing steps and after electrophoresis with continuous electroelution ((e) and (f)).

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