Figure 1.

Extracellular E^rns R171 is a monomer. (a) During the Strep-tag purification process, each sample was collected and equal volumes of the HEK cell supernatant (SN), the five wash steps, the six elution steps as well as the flow through (FT) were separated by SDS-PAGE and stained with Coomassie Brilliant Blue. Purification of the wild-type (C171) and monomeric mutant (R171) are depicted on the left and right panel, respectively. The molecular weights of the proteins are indicated on the left according to the BenchMark™ pre-stained protein ladder. (b) Purified, soluble E^rns C171 and R171 (2.5 µg each) of the BVDV strain Ncp7 were incubated with or without β-mercaptoethanol (2-ME) as indicated. Samples were analysed by Western blot using an anti-E^rns antibody for the non-reducing condition (left panel) or an anti-Strep tag antibody for the reducing condition (right panel). PageRuler Plus pre-stained protein ladder was used for size determination. The signal was detected using a CCD camera with an exposure time of 10 s (left panel) or 5 s (right panel). One representative experiment out of three is shown.