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. 2015 Jan 13;14:3. doi: 10.1186/s12934-014-0186-0

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© Lauber et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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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.