Figure 7.

Immunoisolation of E1-G_CT containing membranes. BHK-E1-G_CT cells were homogenized and separated into total homogenate (TH), postnuclear supernatant (PNS), nuclear pellet (NP), smooth membranes (SM), rough membranes (RM), and immunoisolate fractions (bound) prepared using P5D4-coated magnetic beads. Equivalent volumes normalized to starting material for each fraction were electrophoresed through 10% polyacrylamide gels followed by transfer to PVDF membranes. E1-G_CT was detected by probing membranes with rabbit antibody directed against the CT domain of VSV G followed by goat anti-rabbit IgG-HRP and ECL detection. The smooth membrane fraction was used as the starting material for immunoisolation. Most of the E1-G_CT was recovered in the bound fraction; however, a significant portion did not bind to the beads (unbound).