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. Author manuscript; available in PMC: 2011 Feb 24.
Published in final edited form as: Cell Immunol. 2010 Feb 24;263(1):41–48. doi: 10.1016/j.cellimm.2010.02.014

Figure 3. Fibrinogen does not diminish antigen-antibody binding.

Figure 3

Fibrinogen has no discernible effect on binding strength, but high levels of fibrinogen (5 mg/ml) results in increased total binding of fibrinogen to immobilized anti-GFP IgG. Experimental details: GFP was immobilized onto high binding polystyrene microplates, blocked with BSA, and incubated with varying amounts of biotinylated antibody for 1 hour at 25°C in the presence or absence of fibrinogen (0, 1 or 5 mg/ml). To assess unspecific binding, excess GFP was added as competitor. Bound ligand was detected using a colorimetric Streptavidin-Horseradish peroxidase system, and specific binding determined as the difference between bound antibody in the absence of GFP and bound antibody in the presence of GFP. Binding data was analyzed with GraphPad Prism using non-linear curve fitting and the one binding site model. Experiment was performed three times with similar results.