FIG. 2.
Demonstration of functional receptors for Stx2 in the neutral glycolipid fraction of mouse brain. Standard mixtures of neutral glycolipids (lane A) and neutral glycolipids of the mouse brain (lane B) were separated by TLC and visualized with orcinol reagent. Glycolipids cross-reacting with the epitope of Gb3 were identified by immunostaining on TLC-blotted PVDF membranes (lane C from the blot in lane A; lane D from the blot in lane B) with an anti-Pk MAb, followed by treatment with peroxidase-conjugated goat anti-mouse IgM F(ab′)2 and DAB-H2O2. The TLC-blotted membranes used for lane D was incubated with 1 μg of Stx2 per ml; and glycolipids to which Stx2 bound were visualized by immunostaining with an anti-Stx2 MAb, followed by treatment with biotinylated anti-mouse IgG F(ab′)2, avidin-peroxidase complex, and DAB-H2O2 (lane E).