Protein purification was
carried by the affinity chromatography protocol previous described
by Ramos and co-workers [47] using Sephadex G-50 column
(10 50 cm). The ConM was complexed with IAA in presence of
light for 30 min at 310 K and haemagglutinating activity (HA) was
done to confirming that indole binding does not alter the carbohydrate
recognition.
Subsequently, 200 mL suspension of 2% rabbit erythrocytes suspension
containing 0.15 M NaCl was added to each well. HA was
measured after 30 min of incubation at 310 K and 30 min of incubation
at room temperature.