Fig.3.
The TR–FRET assay follows competitive inhibition mediated by IgE-Fc and FcεRIα-specific ligands. (A) Inhibition with FcεRIα lacking a His tag. Here, 25 nM AF488-Fc premixed with increasing concentrations of FcεRIα lacking a His tag (0– 200 nM) was tested in the presence of 15 nM FcεRIα and 5 nM Tb-Ab. (B) Inhibition with anti-IgE omalizumab (Xolair). Here, 25 nM AF488-Fc premixed with increasing concentrations of Xolair (0–200 nM) was tested in the presence of 15 nM FcεRIα and 5 nM Tb-Ab. (C) Inhibition with the IgE-Fc ligand DARPin E2_79. Here, 25 nM AF488-Fc premixed with increasing concentrations of E2_79 (0–200 nM) was tested in the presence of 15 nM FcεRIα and 5 nM Tb-Ab. (D) Inhibition with anti-FcεRIα Mab15-1. Here, 25 nM AF488-Fc was tested in the presence of 15 nM FcεRIα and 5 nM Tb-Ab premixed with increasing concentrations of mAb15.1 (0–200 nM).