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. 2016 Dec 13;7:567. doi: 10.3389/fimmu.2016.00567

Figure 4.

Figure 4

ELISA to assess interaction between gC1qR and gh substitution mutants. Microtiter wells were coated with different quantities (0.25, 0.5, and 1 μg/well) of gC1qR. After blocking and washing, the wells were incubated with 2.5 μg/well of (A) ghA, R162A and R162E; (B) ghB, R114A, R114Q, R163A, R163E, R129A, R129E, T175L, and H117D; and (C) ghC, for 90 min at 37°C and 90 min at 4°C. Bound proteins were detected with anti-MBP monoclonal antibody followed by goat anti-mouse IgG–HRP conjugate. Data are representative of three experiments. (D) Ligand blot to show binding of ghA mutants R162A and R162E to gC1qR: PVDF membrane strips containing gC1qR were reacted with ghA, R162A and R162E, and then probed with anti-MBP monoclonal antibody followed by goat anti-mouse IgG–HRP.