American Society of Hematology

Fig. 2.

$Fig. 2. Binding of 125I–SDF-1α to Fn in solution as determined by EMSA shows a Kd of approximately 10 nmol/L. / (A) Indicated concentrations of 125I–SDF-1α were incubated in solution with 200 nmol/L Fn, then run on a native 4% to 20% PAGE gel. Duplicate lanes represent independent binding assays performed simultaneously. Position of free SDF-1α and Fn are indicated (both high-molecular-weight bands are positive in Fn Western blot; the lower of the 2 comigrates with myosin, approximately 250 kd). (B) Similar gels were used to measure the amount of125I–SDF-1α in the bound and free fractions, as described in the text, and were plotted in a Scatchard analysis. Inset: typical standard curve used to determine the bound and free fractions. (C) Binding to other ECM proteins was examined by EMSA. Lane 1, Fn; lane 2, collagen IV; lane 3, laminin 1; lane 4, laminin 2; lane 5, no ECM protein (125I–SDF-1α alone).$

Binding of ¹²⁵I–SDF-1α to Fn in solution as determined by EMSA shows a K_d of approximately 10 nmol/L.

(A) Indicated concentrations of ¹²⁵I–SDF-1α were incubated in solution with 200 nmol/L Fn, then run on a native 4% to 20% PAGE gel. Duplicate lanes represent independent binding assays performed simultaneously. Position of free SDF-1α and Fn are indicated (both high-molecular-weight bands are positive in Fn Western blot; the lower of the 2 comigrates with myosin, approximately 250 kd). (B) Similar gels were used to measure the amount of¹²⁵I–SDF-1α in the bound and free fractions, as described in the text, and were plotted in a Scatchard analysis. Inset: typical standard curve used to determine the bound and free fractions. (C) Binding to other ECM proteins was examined by EMSA. Lane 1, Fn; lane 2, collagen IV; lane 3, laminin 1; lane 4, laminin 2; lane 5, no ECM protein (¹²⁵I–SDF-1α alone).

This Feature Is Available To Subscribers Only