Journal: PLoS ONE
Article Title: A New Ligand-Based Method for Purifying Active Human Plasma-Derived Ficolin-3 Complexes Supports the Phenomenon of Crosstalk between Pattern-Recognition Molecules and Immunoglobulins
doi: 10.1371/journal.pone.0156691
Figure Lengend Snippet: Analysis of ficolin-3 preparation (1 μg of final product) fractionated on a 10% polyacrylamide gel under reducing (lanes 1, 2, 5–8) or non-reducing (lanes 3–4) conditions. Coomassie Blue stain (lanes 1, 3) and Western blotting using anti-ficolin-3 (lanes 2, 4), anti-MASP-1 (lane 5), anti-MASP-2 (lane 6), and anti-MASP-3 (lane 7) antibodies. The last lane (8) is the strip 4 re-probed with anti-human IgG, IgA and IgM. MASP bands are marked with asterisks (*). A, b, c–heavy chains of IgM (a), IgA (b), IgG (c); d- ficolin-3 monomer; e- light chains of Ig. The positions of the molecular mass markers are indicated in kDa.
Article Snippet: After blocking for 2 h with 5% skim milk, membranes were incubated with goat polyclonal anti-MASP-1 (catalogue number: sc-50841, Santa Cruz Biotechnology), rat monoclonal anti-human MASP-2/MAp-19 (Hycult Biotech, catalogue number: HM2191), or goat polyclonal anti-complement MASP-3 (catalogue number: AF1724-SP, R&D Systems) for 2 h at room temperature.
Techniques: Staining, Western Blot, Stripping Membranes