TechNotes No. 12-2000

Binding of a plant lectin horseradish peroxidase conjugate to a-D-galactoside GlycoWell™ plates.

Introduction
Binding of lectins to immobilized saccharides and subsequent inhibition of such a binding event is often used for evaluating lectin inhibitors. The use of GlycoWell™ plates facilitates an ELISA assay involving the binding of lectins to the wells. This TechNote describes the binding of an a-D-galactopyranoside recognizing lectin Maclura pomifera1, to a-D-galactopyranoside GlycoWell™ plates.

Experimental

  1. Lectin incubation. Add 100 L lectin solution to each well. Leave for 45 min
  2. Washing. Empty the wells and wash three times with CovaBuffer. Leave the last wash in the wells for 15 min. Wash with citrate/phosphate buffer once.
  3. Substrate reaction. Add 100 L substrate solution to each well. Read optical density at 490 nm in a microwell plate reader.
Discussion
The results demonstrate that a a-D-galactopyranoside GlycoWell™ plate efficiently binds a a-D-galactopyranoside-recognizing plant lectin, Maclura pomifera (Figure 1). Thus, GlycoWell plates are suitable for detecting carbohydrate binding properties of proteins.

Materials
Figure 1. A Maclura pomifera-horseradish peroxidase conjugate binds to a-D-galactopyranoside GlycoWell™ plates (SW-01-007), but not to a GlycoWell™ plate devoid of saccharide (SW-00-001).

Reference.
1. Jones, J.M. and Feldman, J.D. J. Immunol., 1973, 3, 1765.