Figure 4.
SDS-PAGE analysis of recombinant proteins purified by Ni-chelating chromatography and used in the solid-phase binding assays. Culture media from HEK-293T cells transiently transfected with cDNA constructs encoding the different recombinant proteins were fractionated by Ni-chelating HPLC as described.16 Aliquots of the isolated proteins were fractionated by SDS-PAGE (10% acrylamide). After electrophoresis, proteins were either transferred to nitrocellulose membranes and detected by chemiluminiscence using anti-myc or anti-HA antibodies (1:500)12,14 (A) or detected by silver nitrate staining (B).