TechNotes No. 9-2000

Binding of a plant lectin — horseradish peroxidase conjugate to 2–acetamido–2–deoxy–bD–galactopyranoside GlycoWell™ plates.

Binding of lectins to immobilized surfaces 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 2–acetamido–2–deoxy–bD–galactopyranoside recognizing lectin, Ricinus communis toxin RCA 1201, to 2–acetamido–2–deoxy–bD–galacto-pyranoside 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. Leave for 15 min. Add 100 mL 1M H2SO4(aq) to each well and read optical density at 490 nm in a microwell plate reader.
The results demonstrate that a 2–acetamido–2–deoxy–bD–galacto-pyranoside GlycoWell™ plate efficiently binds a 2–acetamido–2–deoxy–bD–galactopyranoside-recognizing plant lectin, RCA 120 (Figure 1). Thus, GlycoWell plates are suitable for detecting carbohydrate binding properties of proteins.


Figure 1. A RCA 120-horseradish peroxidase conjugate binds to 2–acetamido–2–deoxy–bD–galacto-pyranoside GlycoWell™ plates (SW-01-002), 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.