3.5.2. Western blot analysis of sperm-extracted protein using polyclonal antibodies anti-acrosin I and gelatinases of sperm extract
When NPGB was present three main bands were observed with molecular weights ranging from 35 to 66 kDa in fresh semen and fresh semen containing DMA. After freezing-thawing, a band with a molec- ular weight of 35 kDa disappeared and two new bands with molecular masses of 36.5 and 38.7 kDa appeared (Fig. 9A).
When NPGB was absent three bands were observed with molecular weights ranging from 30 to 41 kDa in fresh semen and fresh semen containing DMA. The bands corresponding to acrosin I and II were ob- served. After freezing-thawing, two additional bands with molecular weights of 24 and 19 kDa were ob- served (Fig. 9B).
Using gelatin SDS-PAGE three bands were ob- served with the molecular weight ranging from 30 to 41 kDa in fresh semen. In fresh semen containing DMA one additional band with a molecular weight of 32 kDa was observed, and after freezing-thawing its intensity increased. No bands were observed below the band with a molecular weight of 30 kDa (Fig. 9C).
3.5.3. Seminal plasma amidase activity and Western blot analysis of seminal plasma protein
No differences were found in amidase activity of seminal plasma of semen before and after freezing-