As well as providing this step as an integral part of our service to make a recombinant virus, we can also test clients’ own viruses for expression. We require at least 1 ml of high titre virus (if titre is unknown or unsure, we strongly recommend titrating the virus first by plaque-assay or baculoQUANT).
OET will carry out q quick test for expression by infecting dishes of Sf9 cells with seed stock virus (200 ul); a dish of control cells will also be mock-infected with culture medium. At 48 hours post-infection, the cells and culture medium will be harvested and analysed separately for expression of protein by Western Blot using either anti-tag anti-serum (usually anti-His-tag) or using primary antisera supplied by the client or purchased commercially by OET (on the instructions of, and at cost to, the client). Using the results of these experiments, a strategy for producing and purifying the protein will be made.
Please contact us for further details of this service.
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