FIG. 1.
(A) C3c levels in serum samples measured by a C3c-specific sandwich ELISA. Serum was incubated with inulin, zymosan, aggregated IgG (HA IgG), CVF, or SPS and control serum without application. After incubation, sera were made 10 mM with respect to EDTA, diluted 1:320. The C3c levels were assessed with a C3c-specific MAb (F1-4) as catching antibody and levels were measured as OD490-650 units. Error bars indicate two times the standard deviation of double determinations. (B) Western blot of human serum activated with inulin, zymosan (Zymo), aggregated IgG (HA IgG), CVF, or SPS and control serum without application. Nonreducing SDS-PAGE of serum samples diluted 1:40. C3 bands were visualized with the anti-human C3 monoclonal antibody (HAV3-4). We observed three bands with apparent molecular sizes of 180, 175, and 140 kDa corresponding to native C3, C3b/iC3b, and C3c, respectively. The MW marker was Precision Plus Protein Prestained (Bio-Rad).