Assay Report Card

ID: ALA3706058

Type: Binding

Description: Enzyme Assay: Virus propagation: Influenza A/Brisbane/59/2007 (H1N1) virus was propagated in Madin-Darby Canine Kidney (MDCK) cells. Seed virus was inoculated into 10 confluent 75 cm2 monolayers which were incubated at 37° C. for 2-3 days and harvested when full cytopathic effect was observed. Virus purification: The pooled cell lysates were frozen and thawed and clarified by centrifugation at 3000 g for 20 mM. The supernatant was subjected to ultracentrifugation at 25,000 rpm for 90 min and the virus containing pellet was resuspended in 2 mL of MegaVir medium (Hyclone). Virus concentration was assessed by hemagglutination (HA) at 40,960 HA units. Virus inactivation: NP40 (Fluka) was added to the purified influenza virus at a final concentration of 0.2% and the mixture was incubated at room temperature for a total of 3 hours and at 4° C. for 6 h. The inactivation of the virus was confirmed by the Tissue Culture Infectious Dose assay.