Figure 7.

Lectin blot analysis of affinity-purified gelatin-binding proteins from human seminal plasma. Affinity-purified gelatin-binding proteins from human seminal plasma were resolved by 12.5% SDS-PAGE under reducing conditions, blotted onto Immobilon-P–PVDF membrane and probed with biotinylated lectins: RCA I (Ricinus communis agglutinin I), ConA (lectin from Canavalia ensiformis), LCA (Lens culinaris agglutinin), SNA (Sambucus nigra agglutinin) and MAA (Maackia amurensis agglutinin). Bound lectins were visualized using a mixture of avidin and biotinylated-HRPO followed by addition of H₂O₂/3,3′-diaminobenzidine. Numbers indicate molecular mass (kDa). Lectin binding to higher molecular mass species was more noticeable when the sample was resolved on 20% gel (upper panel) because of the generally known diffuse appearance of glycoproteins on SDS-PAGE. Asterisks indicate main lectin-reactive bands.