Mouse anti his tag

To verify protein expression, the harvested Sf-9 cells infected with recombinant AcMNPV at 72 h post-infection were subjected to SDS-PAGE analysis and Western blotting. Uninfected Sf-9 cells, which were similarly treated, were used as the negative control. Western blotting was performed with a mouse anti-His-tag (1:1000 diluted with TBST; Abcam, Cambridge, UK) overnight at 4 °C. The secondary antibody was alkaline phosphatase-conjugated goat anti-mouse IgG (1:5000 diluted with TBST; Thermo Fisher, Waltham, MA, USA) and visualized using the enhanced chemiluminescence method.

To confirm that the rVP35-VP4 protein can be expressed in BmE cells, the BmE cells were collected 72 h after infection with BmNPV-VP35-VP4 for SDS-PAGE and Western blot analysis. BmE cells infected with wild-type BmNPV were used as a negative control. The cell extracts were run on 12% SDS-PAGE and the separated proteins were transferred onto PVDF membranes. After blocking with 5% skim milk in phosphate buffered saline (PBS)–Tween (PBST) for 2 h at 37 °C, the membrane was incubated with a mouse anti-His-tag (1:1000 diluted with TBST, Abcam, Cambridge, UK) and the secondary antibody alkaline phosphatase-conjugated goat anti-mouse IgG (1:5000 diluted with TBST, Thermo Fisher, Waltham, MA, USA) and finally visualized by the enhanced chemiluminescence (ECL) method.