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.
aliquot of supernatant was added to the first well of the column.
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.