Journal: Journal of leukocyte biology

Article Title: Subversion of complement activation at the bacterial surface promotes serum resistance and opsonophagocytosis of Francisella tularensis.

doi: 10.1189/jlb.0807526

Figure Lengend Snippet: Fig. 2. Analysis of the deposition and cleavage of C3 at the surface of SCHU S4. The cleavage profile of C3 eluted from the surface of SCHU S4, pretreated with NHS versus EDTA-plasma, was analyzed by Western blotting using a sheep polyclonal antibody to C3 (A) or a mAb anti-iC3b (B). The inactivation of C3b to iC3b is illustrated by the appear- ance of the 68-kDa 1 and the 41-kDa 2 fragments derived from the ’-chain (110 kDa) of C3b. The 1 fragment is depicted as a band running below the intact -chain of C3 (75 kDa; A). The formation of iC3b was also confirmed by detection of the 68-kDa (1) chain by the specific mAb anti-iC3b (B). The -chain of C3 was not recognized by this mAb. The results presented are representative of three separate experi- ments.

Article Snippet: Sheep anti-human polyclonal antibodies to complement factor C3 were from Quidel Corp.

Techniques: Clinical Proteomics, Western Blot, Derivative Assay

Journal: Journal of leukocyte biology

Article Title: Subversion of complement activation at the bacterial surface promotes serum resistance and opsonophagocytosis of Francisella tularensis.

doi: 10.1189/jlb.0807526

Figure Lengend Snippet: Fig. 1. Deposition of C3 fragments on the surface of LVS and SCHU S4 analyzed by ELISA. Microtiter plate wells coated with the bacteria (1107/well) were incubated with NHS or Mg-EGTA NHS at 37°C for 5, 10, 20, 30, 45, and 60 min. Primary mAb antibodies against the different C3 frag- ments (C3d or iC3b) were added, and bound antibodies were detected with a HRP-con- jugated secondary antibody. (A) Deposition of C3 at the surface of LVS treated with NHS or Mg-EGTA NHS as detected by anti- C3d. (B) Deposition of C3 at the surface of SCHU S4 treated with NHS or Mg-EGTA NHS as detected by anti-C3d. (C) Deposi- tion of iC3 at the surface of LVS treated with NHS or Mg-EGTA NHS as detected by anti-iC3b. (D) Deposition of C3 at the sur- face of SCHU S4 treated with NHS or Mg- EGTA NHS as detected by anti-iC3b. Val- ues are ODs measured at 405 nm. The OD values of the controls incubated with serum and secondary antibodies only were sub- tracted from the values shown. The results are from three separate experiments. Statis- tically significant differences between un- treated NHS and corresponding Mg-EGTA NHS are marked (*, P0.05).

Article Snippet: Sheep anti-human polyclonal antibodies to complement factor C3 were from Quidel Corp.

Techniques: Enzyme-linked Immunosorbent Assay, Bacteria, Incubation