<p>(<b>A</b>) Recombinant GST VWF-A2 protein (100 nM) was mixed with recombinant full-length or truncated fH (30 nM) and precipitated using glutathione agarose beads. The precipitated proteins were immunoblotted (IB) with anti-fH antibody for examining the association between VWF-A2 and fH molecules. Immunoblotting with anti-VWF antibody served as loading control. A representative blot is shown (n=3). (<b>B</b>) We repeated the above experiment but used a monoclonal anti-fH antibody (clone 90X) recognizing SCR1 to immunoblot fH fragments precipitated with GST-VWF-A2. (<b>C</b>) Recombinant VWF-A1 protein (100 nM) was mixed with recombinant full-length or truncated fH (30 nM) and precipitated using polyclonal anti-VWF antibody. The precipitated proteins were immunoblotted with monoclonal anti-fH antibody (clone 90X) for examining the association between VWF-A1 and fH molecules. (<b>D</b>) SPR sensograms were generated by injecting recombinant fH proteins (F1, F2, F3, or F4) at a concentration of 10 µg/ml over immobilized VWF-A2 or VWF for 60s followed by dissociation for 2 min. (<b>E</b>) SPR sensograms generated by injecting fH proteins (F1, F2, F3, and F4) over immobilized ovalbumin (OVA), served as a negative control.</p
