igg fraction of sheep polyclonal antibodies against tfpi  (Haematologic Technologies)

Journal: PLoS ONE

Article Title: Tissue Factor Pathway Inhibitor 2 Is Found in Skin and Its C-Terminal Region Encodes for Antibacterial Activity

doi: 10.1371/journal.pone.0052772

Figure Lengend Snippet: ( A ) Fibrin clots obtained from patients with chronic wounds were analyzed by immunoblotting using polyclonal antibodies against the C-terminal of TFPI-2. Citrate plasma (CP) and recTFPI-2 were used as negative and positive controls. ( B ) TFPI-2 C-terminal epitopes are found in human wounds. Gold-labeled antibodies against the C-terminal of TFPI-2 (visualized as black dots) show that this epitope is found on bacteria present in fibrin slough from a P. aeruginosa infected chronic wound. ( C ) Proteolytic cleavage of recTFPI-2 with human leukocyte elastase (HLE). Left panel: Western blot analysis of degraded protein using polyclonal antibodies against the C-terminal part of TFPI-2. Right panel: Coomassie stained gel of the same material. The arrow indicates the sequenced fragment. ( D ) Digested C-terminal TFPI-2 fragment binds to bacterial surface. Intact and cleaved recTFPI-2 was incubated with E. coli for 1 h at 37°C and supernatants (S) and bacterial bound proteins were (pellet; P) analyzed by western blot.

Article Snippet: This blocking procedure was followed by overnight incubation at 4°C with 1 μg/ml of polyclonal rabbit antibodies raised against the C-terminal TFPI-2 sequence CAKALKKKKKMPKLRFASRIRKIRKKQF (CAK28 antibodies) (Innovagen AB).

Techniques: Western Blot, Clinical Proteomics, Labeling, Bacteria, Infection, Staining, Incubation