Carbohydrate specific identification and purification of Eutirucallin.
2014-02-18T04:00:01Z (GMT) by
<p><b>A.</b> Binding assay protein-carbohydrate with agarose beads adsorbed with different carbohydrates. “Beads” impregnated with different carbohydrates were incubated with the protein extract, later eluted from the resin after addition of 2-mercaptoethanol (2ME+) followed by boiling; supernatants were evaluate4d by SDS-PAGE. Lactose, mannose, N-acetyl-D-galactosamine (GalNAc) and N-acetylglucosamine (GlcNAc) correspond to the binding proteins for the corresponding sugars. <b>B.</b> Lectin purification by affinity chromatography. Approximately 2.41 mg of total protein was applied in 2 mL of α-D-Lactose column previously balanced with PBS, and eluted with galactose (0.4 M). The fractions collected during the purification were spectrophotometrically monitored (A<sub>280nm</sub>). <b>C.</b> Evaluation of proteins of the protein extract (PE) and Eutirucallin in bland and highly denaturating conditions. 2ME+ and 2ME- correspond, respectively, to the presence and absence of 2-mercaptoethanol. Mr, corresponds to the molecular pattern, whose numbers on the left indicate the mass values.</p>