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. Author manuscript; available in PMC: 2013 Dec 1.
Published in final edited form as: Clin Immunol. 2012 Sep 28;145(3):241–250. doi: 10.1016/j.clim.2012.09.007

Figure 5.

Figure 5

Stabilizing activity for the classical pathway C3 convertase in the patient’s serum is lost by removal of IgG. Serum IgG was removed by one or two adsorption(s) with Protein G Sepharose. (A) The effect of IgG depleted serum on C3 convertase stability was assessed in an hemolysis assay monitoring convertase decay as described in Methods. Samples were allowed to decay for 30 min at 30°C. Shown is the mean +/−SEM of 3 experiments. Removal of IgG from the patient’s serum (bars 4 and 5) partially abrogated hemolysis mediated by the stabilized convertase whereas there was no effect on IgG removal from NHS (bars 7 and 8). Decay of the convertase in the presence of NHS and IgG depleted patient serum was accelerated compared to the buffer alone control due to the presence of C4BP in the serum. (B) Western blot assessment of the efficacy of IgG removal from sera by Protein G adsorption. The blots were performed as described in Methods and developed using donkey anti-human IgG (1:5,000).