FIG. 5.

Purification of IMTGP-2 by chromatography on Jacalin agarose. (a) K88ac adhesin binding activity (□) was determined with biotinylated K88ac adhesin as described in Materials and Methods. Protein concentration was monitored by UV absorption at 280 nm (•). Horizontal lines represent the pooled fractions containing K88ac adhesin receptors. Fraction 4 was obtained by eluting the column with buffer B containing 0.3 M melibiose. Fraction 5 was obtained after elution with buffer B containing 0.3 M methyl-α-d-galactopyranoside (MeGal). (b) Glycoproteins (25 μg) were separated by SDS-PAGE (7% polyacrylamide) and transferred to PVDF membranes. (A) K88ac-binding activity was detected with biotinylated K88ac adhesin. (B) Glycoproteins were detected by PAS staining. (C) Proteins were detected by Coomassie blue staining. Lane 1, biotinylated molecular mass markers; lanes 2 and 3, Jacalin fractions 4 and 5, respectively.