Aec hrp substrate kit

The LCMV stock was prepared and titrated according to the previous study [31 (link)]. Briefly, BHK cells were infected with virus for 72 h. The culture supernatant was collected and centrifuged (350 g, 10 min, 4°C) to remove cell debris. Viral stock was stored at −80°C for future use. For titration of virus, Vero cells were infected with a series of 10-fold viral dilutions for 90 min, followed by a methylcellulose overlay. After 4 days of culture, cells were washed and incubated with mouse anti-LCMV polyclonal Ab (Fitzgerald, Acton, MA), followed by incubation with peroxidase (HRP)-conjugated anti-mouse IgG (Southern Biotech, Birmingham, AL). The AEC HRP Substrate Kit (Enzo Life Sciences, Farmingdale, NY) was used for immunocytochemical procedures. Viral titres were calculated by counting the numbers of positive clusters.

The LCMV stocks were prepared and titrated according to a modified method. Briefly, virus was incubated with baby hamster kidney (BHK) cells for 72 h. The culture fluid was centrifuged for 10 min at 350 g, 4°C, and stored at −70°C. For quantitation of the virus, Vero cells were cultured with a series of 10-fold virus dilutions for 90 min, followed by a methylcellulose overlay. After 4 days of culture, cells were first incubated with mouse anti-LCMV polyclonal antibody (Fitzgerald, Acton, MA), followed by incubation with Peroxidase (HRP)-conjugated anti-mouse IgG (Southern Biotech, Birmingham, AL). The AEC HRP Substrate Kit (Enzo life Sciences, Farmingdale, NY) was used for immunocytochemical procedures. Viral titers were calculated by counting the numbers of positive clusters.