Goat anti hamster igg fc

Serum samples were inactivated with γ-irradiation (2 mRad) and analyzed as previously described [63 ]. In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house) per well and incubated overnight at 4°C. After blocking with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT), serially diluted 2-fold serum samples (duplicate, in blocking buffer) were incubated for 1 h at RT. Spike-specific antibodies were detected with goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) for 1 h at RT and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120–0047). The reaction was stopped with KPL stop solution (Seracare) and read at 450 nm. Plates were washed 3 to 5 × with PBS-T (0.1% Tween) for each wash. The threshold for positivity was calculated as the average plus 3 × the standard deviation of negative control hamster sera.

Serum samples were inactivated with γ-irradiation (2 mRad) and analyzed as previously described [28 (link)]. In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house) per well and incubated overnight at 4 °C. After blocking with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT), serially diluted 2-fold serum samples (duplicate, in blocking buffer) were incubated for 1 h at RT. Spike-specific antibodies were detected with goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) for 1 hr at RT and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120-0047, Milford, MA, USA). The reaction was stopped with KPL stop solution (Seracare) and read at 450 nm. Plates were washed 3 to 5× with PBS-T (0.1% Tween) for each wash. The threshold for positivity was calculated as the average plus 3× the standard deviation of negative control hamster sera.

Serum samples were inactivated with γ-irradiation (2 mRad) and analyzed as previously described (Yinda et al., 2020 ). In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house) per well and incubated overnight at 4 °C. After blocking with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT), serially diluted 2-fold serum samples (duplicate, in blocking buffer) were incubated for 1 h at RT. Spike-specific antibodies were detected with goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) for 1 h at RT and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120–0047). The reaction was stopped with KPL stop solution (Seracare) and read at 450 nm. Plates were washed 3 to 5 × with PBS-T (0.1 % Tween) for each wash. The threshold for positivity was calculated as the average plus 3 × the standard deviation of negative control hamster sera.