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. 2009 Mar 18;96(6):2313–2320. doi: 10.1016/j.bpj.2008.12.3900

Figure 5.

Figure 5

Shear induced conformation changes in VWF detected using bis-ANS. (A) Representative spectra for an experiment performed with 60 μg/mL VWF. Bis-ANS binding to VWF increases absolute fluorescence (F(G)(λ), counts per second) by 5–10-fold over the HEPES buffer signal (squares). Application of shear for 5 min at 6000 or 9600/s further increased the absolute bis-ANS signal by 110–220%, compared to results obtained with no-shear control (G = 0/s). (B) Shearing 50 μg/mL BSA does not change signal due to bis-ANS BSA interaction. (C) Concentration normalized spectra (Fc(G)(λ)) for VWF. Raw data are from panel A. (D) Comparison of Fc(G)max/Fc(G = 0/s)max for VWF under different shear conditions. Data in panels A–C are representative of eight independent experiments. Results in panel D are presented as mean ± SE (N ≥ 3 at each shear rate). ∗p < 0.05 with respect to shear rate = 30/s.