Hrp conjugated mouse anti human his antibody

For detection of direct interactions between Ebola virus GPs and ADI-15946, His-tagged GPs 2 μg/ml were first coated with coating buffer (0.1 M Na₂CO₃ and NaHCO₃ [pH 9.6]) on 96-well plates overnight at 4°C. Wells were washed with PBST (PBS plus 0.2% Tween 20) and saturated for 1 h at 37°C with PBS–4% nonfat milk. Three-fold dilutions of ADI-15946 were added, and the mixture was incubated for 1 h at 37°C. Bound particles were detected with HRP-conjugated goat anti-human IgG antibody (Invitrogen; A18817). PS-associated ELISAs were carried out as previously described (57 (link)). PS (Sigma; P7769) and phosphatidylcholine (PC) (Sigma; P3556) were first dissolved in chloroform as a stock solution. Then, the solution was diluted to 5 μg/mL with methanol and added to the ELISA plate. Upon being dried, the plates were blocked with Tris-buffered saline (TBS)–4% bovine serum albumin (BSA) fraction V. TIM-1 ECD, TIM-1 IgV, TIM-1 ΔIgV, and control protein were diluted in TBS–10 mM CaCl₂, and the mixture was added to the wells and incubated at 37°C for 1 h. The plates were washed with TBST (TBS plus 0.05% Tween 20) before incubation with HRP-conjugated mouse anti-human His antibody (Biolegend; 652503). The absorbance at 450 nm was measured by microplate reader (Thermo Fisher Scientific).

For detection of direct interactions between Ebola virus GPs and ADI-15946, His-tagged GPs 2 μg/ml were first coated with coating buffer (0.1 M Na₂CO₃ and NaHCO₃ [pH 9.6]) on 96-well plates overnight at 4°C. Wells were washed with PBST (PBS plus 0.2% Tween 20) and saturated for 1 h at 37°C with PBS–4% nonfat milk. Three-fold dilutions of ADI-15946 were added, and the mixture was incubated for 1 h at 37°C. Bound particles were detected with HRP-conjugated goat anti-human IgG antibody (Invitrogen; A18817). PS-associated ELISAs were carried out as previously described (57 (link)). PS (Sigma; P7769) and phosphatidylcholine (PC) (Sigma; P3556) were first dissolved in chloroform as a stock solution. Then, the solution was diluted to 5 μg/mL with methanol and added to the ELISA plate. Upon being dried, the plates were blocked with Tris-buffered saline (TBS)–4% bovine serum albumin (BSA) fraction V. TIM-1 ECD, TIM-1 IgV, TIM-1 ΔIgV, and control protein were diluted in TBS–10 mM CaCl₂, and the mixture was added to the wells and incubated at 37°C for 1 h. The plates were washed with TBST (TBS plus 0.05% Tween 20) before incubation with HRP-conjugated mouse anti-human His antibody (Biolegend; 652503). The absorbance at 450 nm was measured by microplate reader (Thermo Fisher Scientific).