Figure 7.
Specificity of a LYVE-1 receptor polyclonal serum. The specificity of an affinity-purified rabbit polyclonal antiserum generated against soluble LYVE-1 receptor and its capacity to block HA binding were assessed in microtiter plate binding assays. In the top panel, wells coated with either LYVE-1 Fc (filled circles), the CD44H ectodomain fragment CD44158his (triangles), or the control fusion protein ICAM-2 Fc (squares) were incubated with appropriately diluted LYVE-1 specific polyclonal antiserum, and binding was detected with peroxidase-conjugated anti–human Ig (see Materials and Methods). As a control, a second set of LYVE-1–coated wells was reacted with appropriately diluted preimmune serum (open circles). In the bottom panel, wells coated with LYVE-1 Fc were incubated with soluble biotinylated HA (5 μg/ml), in the presence of increasing concentrations of either LYVE-1 antiserum or control preimmune serum followed by detection of bound HA as described in Materials and Methods. Data in each case are the mean ± SEM of triplicate determinations.