Editing Expression of HA NA using Baculovoirus (section)

=== <br>Harvest of cells and confirmation of protein expression<br> ===

<br> 1. Harvest cells 6 days post transfection by centrifugation at 2,000 x g for 5 min at room temperature. The supernatant contains the recombinant baculovirus - this will be used to generate working stocks right away, or can be stored at 4 °C for years.<br>

<br> 2. To check protein expression collect pellets and resuspend in 500 μl of PBS. Mix 50 μl of the suspension with 50 μl of SDS-PAGE loading buffer (containing a reducing agent). Run SDS-PAGE and perform a Western blot analysis with an anti-hexahistidine antibody to confirm gene expression. Include cells from the mock transfection as negative controls.<br>