Figure 2
From: Expression of the functional recombinant human glycosyltransferase GalNAcT2 in Escherichia coli
Purification of HisDapGalNAcT2. SDS-PAGE (A) and immunoblot analyses (B and C) of flow-through, wash and elution fractions collected during Ni2+-NTA purification (see Materials and methods) of soluble HisDapGalNAcT2. Insoluble HisDapGalNAcT2 (PC1) produced in Origami™ 2(DE3)pLysS and verified by ESI-MS or His-Em-GFP (PC2, 27 kDa) are included as controls. (A) Coomassie staining detected protein bands with an expected molecular weight (MW) of ca. 61.7 kDa in lanes 7–12. HisDapGalNAcT2 (arrows) was identified by immunoblotting using (B) specific polyclonal anti human GALNT2 antibody or (C) penta-His antibody against the N-terminal His-Tag.