TechNotes No. 3-1999

Binding of plant lectin — horseradish peroxidase conjugate to b–lactoside GlycoWell™ plates.

Binding of lectins to immobilized saccherides 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 three b–lactose recognizing lectins, Erythrina cristagalli lectin1, peanut agglutinin2, and Ricinus communis toxin RCA 1203, to b–lactoside 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 b–lactoside GlycoWell™ plate efficiently binds different b–lactose–recognizing plant lectins, (Figure 3). Thus, GlycoWell™ plates are suitable for detecting carbohydrate binding properties of proteins.


Figure 1: Three different b–lactoside–recognizing lectin–horseradish peroxidase conjugates bind to b–D–lactopyranoside GlycoWell™ plates (SW-02-001), but not to a GlycoWell™ plate devoid of saccharide (SW-00-001).

1. Iglesias, J.L., Eur. J. Biochem., 1982, 123, 247.
2. Bird, G.W.G., Vox Sang., 1964, 9, 748.
3. Nicolson, G.L. and Blaustein, J. Biochim. Biophys. Acta, 1972, 266, 543.