Western blot of FALP with antifibrinogen antibody.

(A) FALP purified from normal human plasma was run on SDS-PAGE in nonreducing conditions and subjected to Western blot analysis with polyclonal antifibrinogen antibody (goat antifibrinogen IgG, Incstar). Bands defined previously as tp85 and p50 were stained. (B) HiPAC-Aldehyde column chromatography, the first step purification of FALP, was performed in the presence (right panel) and absence (left panel) of 150 mmol/L EACA. Eluates from these preparations (HE) were run on SDS-PAGE under nonreducing conditions and analyzed by Western blot analysis with goat antifibrinogen IgG. Fragments X, Y, and D are marked based on the molecular weight of the bands. (C) FALP prepared in the presence of EACA during HiPAC-Aldehyde (150 mmol/L) and Mono Q (50 mmol/L) column chromatography, were subjected to Western blot analysis as above with goat antifibrinogen IgG. Presence of LBP and FHRP-1 in FALP was confirmed by ELISA and Western blot analysis, respectively. Molecular weight standards are shown in kilodaltons.