Figure 2.
Expression and deglycosylation of ADAM 10 protease (A). After transfection of HEK and HEK APP695 cells with ADAM 10 cDNA, cell lysates were immunoprecipitated with anti-HA antibody, subjected to deglycosylation with N-glycosidase F (PNGase F) (lane 3), or directly analyzed by 4–12% NuPAGE gel system and Western blot. (B) HEK and HEK ADAM 10 cells were surface-biotinylated. After immunoprecipitation with anti-HA antibody (16B12) and elution of the immune complexes, four-fifths of the eluate were incubated with immobilized streptavidin (lanes a), and the remaining one-fifth was directly applied to a 10% SDS/PAGE gel (lanes b) and then blotted onto PVDF membranes.