Coomassie Blue Staining and Western Blot Analysis of the Purified rgD5.

(A) After purification steps, different concentrations of purified rgD5 protein (55 kDa) were separated by SDS-PAGE in a 12% gel and stained overnight with Coomassie Blue R-250. Lane kDa: Unstained Protein MW Marker (Fermentas/Thermo Fisher Scientific); Lane 10: 10 μg rgD5; Lane 5: 5 μg rgD5; Lane 2: 2 μg of rgD5; (B) Western blot was performed with mouse monoclonal antibody (MAb) Anti-6xHis HRP conjugated (Anti-6xHis) or with polyclonal antibodies (PAb) from bovine immunized with inactivated BoHV-5 (Anti-BoHV-5). Lane M: Prestained Low Range Protein MW Standard (Bio-Rad); Lane 1: Purified rgD5; Lane 2: KM71H supernatant of non-transformed yeast cells after methanol induction used as negative control.