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. 2010 Nov 10;5(11):e15443. doi: 10.1371/journal.pone.0015443

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Hein 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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Ficolin-2 or Ficolin-3 was depleted from serum using specific monoclonal antibodies. (A) Depleted serum was applied to SDS-PAGE/Western blotting and probed for Ficolin-2 or Ficolin-3. Lane 1: Ficolin-3 depleted serum. Lane 2: Ficolin-2 depleted serum. Lane 3: Control depleted serum. (B–D) Microtiter plates were coated with acBSA and incubated for 45 min at 37°C with different sera before TCC deposition was detected with a monoclonal antibody. (B) Shows TCC deposition for: NHSP (), Ficolin-2 depleted serum (-♦-), Ficolin-2 depleted serum with addition of 5 µg/ml rFicolin-2 (). (C) Shows TCC deposition for: NHSP (), Ficolin-3 depleted serum (-▪-) and Ficolin-3 depleted serum with addition of 25 µg/ml rFicolin-3 (). (D) Shows TCC deposition for: NHSP (), Ficolin-3 deficient serum −/− (-▴-), Ficolin-3 deficient serum −/− with addition of 25 µg/ml rFicolin-3 (). All graphs show mean ± SD of duplicate wells.

Inline graphic — Ficolin-2 or Ficolin-3 was depleted from serum using specific monoclonal antibodies. (A) Depleted serum was applied to SDS-PAGE/Western blotting and probed for Ficolin-2 or Ficolin-3. Lane 1: Ficolin-3 depleted serum. Lane 2: Ficolin-2 depleted serum. Lane 3: Control depleted serum. (B–D) Microtiter plates were coated with acBSA and incubated for 45 min at 37°C with different sera before TCC deposition was detected with a monoclonal antibody. (B) Shows TCC deposition for: NHSP (), Ficolin-2 depleted serum (-♦-), Ficolin-2 depleted serum with addition of 5 µg/ml rFicolin-2 (). (C) Shows TCC deposition for: NHSP (), Ficolin-3 depleted serum (-▪-) and Ficolin-3 depleted serum with addition of 25 µg/ml rFicolin-3 (). (D) Shows TCC deposition for: NHSP (), Ficolin-3 deficient serum −/− (-▴-), Ficolin-3 deficient serum −/− with addition of 25 µg/ml rFicolin-3 (). All graphs show mean ± SD of duplicate wells.