Goat anti human h l hrp

For detection of HA cross-reactivity, ELISA assays were performed as previously described (Wilson et al., 2014 ). Briefly, Costar Hi Bind plates (Corning Inc., Tewksbury, MA) were coated overnight with the appropriate virus isolate (25 HA units/well) at 4 °C. Plates were blocked for 1 h with PBS/0.1% Tween-20 (PBST) containing 1.5% BSA (blocking buffer) at room temperature. All rmAbs were serially titrated three-fold in blocking buffer and allowed to incubate with antigen-coated plates for 1 h at room temperature. After three PBST washes, wells were probed with goat anti-human (H&L)–HRP (Thermo Scientific) for 1 h at room temperature. Plates were washed three times with PBST and signal was developed with 1 step™ Turbo TMB-ELISA reagent (Thermo Scientific). Reactions were stopped with 1 N sulfuric acid and absorbance was read at 450 nm.

Costar Hi Bind plates (Corning Inc., Tewksbury, MA) were coated overnight with TIV or the individual monovalent inactivated virus constituents at 4 °C. For 2011/12 TIV, 3 μg/ml of total HA was adsorbed, and 1 ug/ml total HA was adsorbed for the individual vaccine constituents. For recHA, 1 ug/ml of respective protein was absorbed. The plates were blocked for 1 h with PBS/0.1 % Tween20 (PBST) containing 1.5 % BSA (blocking buffer) for 1 h at room temperature. All rmAb were serially titrated in blocking buffer and allowed to incubate for 1 h at room temperature. After three PBST washes, the wells were probed with goat anti-human (H&L)-HRP (Thermo Scientific) for 1 h at room temperature. The plates were washed three times with PBST, and the signal was developed with 1 step Turbo TMB-ELISA reagent (Thermo Scientific). The reactions were stopped with 1 N sulfuric acid, and the absorbance was read at 450 nm.