TechNotes No. 2-1999

Binding of a plant lectin — horseradish peroxidase conjugate to bD–galactoside GlycoWell™ plates.

Binding of lectins to immobilized saccharides is an efficient method for evaluting lectin. In addition, competive inhibition lectins1 binding to immobilized saccherides provides a convenient method for screening and characterization of 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 a bD–galactopyranoside recognizing lectin, Ricinus communis toxin RCA 1201, to bD–galactopyranoside GlycoWell™ plates.


  1. Lectin incubation. Add 100 µL 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 µL substrate solution to each well. Read optical density at 490 nm in a microwell plate reader.
The results demonstrate that a bD–galactopyranoside GlycoWell™ plate efficiently binds a bD–galactopyranoside–recognizing plant lectin, RCA120 (Figure 1). Thus, GlycoWell plates are suitable for detecting carbohydrate binding properties of proteins.

Figure 1. A RCA120–horseradish peroxidase conjugate binds to bD–galactopyranoside GlycoWell™ plates (SW-01-006), but not to a GlycoWell™ plate devoid of saccharide (SW-00-001).

1. Nicolson, G.L. and Blaustein, J. Biochim. Biophys. Acta, 1972, 266, 543.