Figure 3.
Signal responses of four ELISAs in two formats (A and B) run on the same plate using HRP-VHH A1 (1:1600 dilution) as the detection antibody. For format A, anti-sEH affinity purified polyclonal antibody (pAb, 3 μg/mL) or antiserum (1:2000 dilution) was used as the capture antibody through passive adsorption on high-binding polystyrene microplate. For format B, nanobody A9 (20 μg/mL) coated in CB or PBS buffer was used as the capture antibody after passive adsorption. The four ELISAs were performed on the same plate using 3% skim milk (SM) for blocking after the coating step. The same reagents involved for different formats are used from the same batch prepared. All same or similar steps were performed at the same time under the same conditions. Error bars indicate standard deviations (n = 3). All coefficient of determination R2>0.99.