Fig. 8.
Determination of the binding affinity of nonasaccharide 9 to AT. (A) The representative autoradiography of the agarose gel in which purified 3-O-[35S]-sulfated nonasaccharide 9 was subjected to electrophoresis through separation zones containing AT at concentrations indicated. Approximately 8000 cpm/lane of nonasaccharide was loaded into each zone. (B) The plot of R/[AT] vs R, where R = (M0 − M)/M0. M0 is the migration of free 35S-labeled nonasaccharide and M is the observed migration of the nonasaccharide in the presence of AT at various concentrations. The plot yields a straight line with a slope of −1/Kd. The linear coefficient (r2) was determined to be 0.97.