Figure 4

SDS-PAGE (A) and Western blot (B) analysis of the proteins present in the culture supernatant of Trichoderma harzianum 1051 grown in liquid medium containing 0.5% casein, for different growing times: 6 h, 24 h, 48 h, and, 72 h. Mw, molecular mass standards. Samples of culture medium containing about 100 μg of proteins were diluted in sample buffer and applied in the gel. Proteins in the SDS-PAGE were silver stained. The nitrocellulose membrane was developed with the polyclonal antibodies raised in mice against the Trichoderma harzianum 1051 purified protease, and alkaline phosphatase-conjugated anti-mouse IgG and the color developing reagents nitro blue tetrazolium and 5-bromo-4-chloro-3-indolyl. The arrows indicate position of the 16, 17, and, 18.8 kDa proteins.