TechNotes No. 4-1999

Binding of a Bandeiraea simplicifolia agglutinin — horseradish peroxidase conjugate to b–Globotrioside GlycoWell™ plates.

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 a aD–galactopyranoside recognizing lectin, Bandeiraea simplicifolia agglutinin (bs–I)1, to b–globotrioside (Gb3) 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 b–globotrioside GlycoWell™ plate efficiently binds a aD–galactopyranoside–recognizing plant lectin, Bandeiraea simplicifolia agglutinin (bs–I) (Figure 1). Thus, GlycoWell™ plates are suitable for detecting carbohydrate binding properties of proteins.


Figure 1. A Bandeiraea simplicifolia agglutinin (bs–I)lectin–horseradish peroxidase conjugate binds GlycoWell™ plates (SW-03-002), but not to a GlycoWell plate devoid of saccharide (SW-00-001).

1. Hayes, C.E. and Goldstein, I.J., J. Biol. Chem., 1974, 249, 1904.