Fig. 2. Ligand blot analysis of the megalin tail binding to MegBP and Dab1. 10 µg of purified GST-MegBP (lanes 1, 4 and 7), GST (lanes 2, 5 and 8) and GST-Dab1 (lanes 3, 6 and 9) were subjected to reducing 10% SDS-PAGE and stained with Coomassie blue (lanes 1-3) or transferred to a nitrocellulose membrane and incubated with an anti-GST antibody (lanes 4-6) or ¹²⁵I-His-megalin tail (lanes 7-9) as described in Materials and Methods. Binding of the ¹²⁵I-His-megalin tail to GST-MegBP and GST-Dab1, but not to GST or GST-MegBP degradation products, was detected by autoradiography.