Table 1. Antibodies used for ELISA assays.
Detected Analyte | Sample | Coating protein | Coating Concentration | Detection Ab | Concentration | Depicted in |
---|---|---|---|---|---|---|
Murine anti-PR8 IgG | Murine sera | PR8 virus | 500 TCID50/mL in 100 mM NaHCO3 | HRP-linked rabbit anti-murine IgG (Sigma; Cat #A9044) | 1:10,000 in 1% BSA PBS-T | Figure 3c |
Ovine IgG in circulation | Murine sera | Purified donkey anti-sheep IgG (Sigma; Cat # SAB3700721) | 5 μg/mL in 100 mM NaHCO3 | HRP-linked anti-sheep IgG (Sigma; Cat # A3415) | 1:15,000 in 1% BAS PBS-T | Figure 4a |
Murine anti-ovine IgG | Murine sera | Ovine pAb | 10 μg/mL in 100 mM NaHCO3 | HRP-linked rabbit anti-murine IgG (Sigma; Cat # A9044) | 1:10,000 in 1% BSA PBS-T | Figure 4b |
Ovine anti-PR8 IgG | Ovine pAb | PR8 virus | 500 TCID50/mL in 100 mM NaHCO3 | HRP-linked anti-sheep IgG (Sigma; Cat # A3415) | 1:15,000 in 1% BAS PBS-T | Figure 5a |
Ovine anti-A/PC IgG | Ovine pAb | A/PC virus | 500 TCID50/mL in 100 mM NaHCO3 | HRP-linked anti-sheep IgG (Sigma; Cat # A3415) | 1:15,000 in 1% BAS PBS-T | Figure 5a |
ELISA protocols were developed to assess the levels of murine anti-PR8 IgG, circulating ovine IgG or murine anti-ovine IgG in sera from infected or treated mice, or to assess the ability of ovine IgG to bind to PR8 and A/PC viruses.