Lectin biotinylated bk 1000 kit

Small intestine sections were assayed with seven lectins (Table 1) (Lectin Biotinylated BK 1000 Kit, Vector Laboratories Inc., Carpinteria, CA, USA) namely: Con A (Concanavalia ensiformis), DBA (Dolichos biflorus), SBA (Glycine max), PNA (Arachis hypogaea), RCA-I (Ricinus communis-I), UEA-I (Ulex europaeus-I) and WGA (Triticum vulgaris) to reveal possible changes of the glycosylation pattern.
In short, paraffin sections were deparaffinized with xylene dehydrated with 100% alcohol twice, 10 min each, and then endogenous peroxidase activity was quenched by incubating 5 min with hydrogen peroxide in methanol 0.3–3.0%.
They were then hydrated, washed in phosphate-buffered saline, and incubated with biotinylated lectins overnight. Then sections were washed with PBS, followed by 10-min incubation with streptavidin-HRP (streptavidin conjugated to horseradish peroxidase in PBS containing stabilizing protein and anti-microbial agents (Vector Laboratories Inc., USA). Finally the bound lectins were visualized by incubation during 4–10 min with a buffered Tris-HCl solution (0.05 M, pH = 6.0) containing 0.02% 3,3′-diamino-benzidine tetrahydrochloride (DAB) and 0.05% H₂O₂ (DAB; Dako, Carpinteria, USA). Positively-stained cells were demonstrated by a dark golden brown coloration. The sections were counterstained with Maeyer haematoxilyn.

To reveal changes of the glycosylation pattern, four small intestine sections of each animal were assayed employing a kit of seven biotinylated lectins (Table 1) (Lectin Biotinylated BK 1000 Kit, Vector Laboratories Inc., Carpinteria, CA, USA). Deparaffinized sections were incubated with the biotinylated lectins overnight as described elsewhere [32 (link)], incubated with Streptavidin-HRP (Vector Laboratories Inc., USA) and bound lectins visualized by 3, 3’-diaminobenzidine (Dako, Carpinteria, USA). Positively-stained cells were revealed by a dark golden brown coloration. The sections were counterstained with Mayer’s haematoxylin.