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. 2009 Mar 24;4(3):e4858. doi: 10.1371/journal.pone.0004858

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Grosskinsky et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Functional activity of CFH was analyzed by measuring factor I-mediated conversion of C3b to iC3b. CFH bound to HcpA coated microtiter plates (A) or to the surface of intact B. recurrentis spirochetes (B) was incubated with C3b and factor I. Reaction mixtures were separated by SDS-PAGE and transferred to nitrocellulose membrane. C3b degradation products were evaluated by detection of α′-chain cleavage fragments of 68, 46 and 43 kDa using biotinylated rabbit anti-C3c IgG followed by peroxidase-conjugated streptavidin. Purified iC3b was included as control.