Journal: The Journal of biological chemistry

Article Title: Mutational analysis of the predicted first transmembrane segment of each homologous half of human P-glycoprotein suggests that they are symmetrically arranged in the membrane.

doi: 10.1074/jbc.271.26.15414

Figure Lengend Snippet: FIG. 2. Endoglycosidase H digestion of wild-type and inactive mutants of P-glycoprotein transiently expressed in HEK 293 cells. Whole cells expressing wild-type or mutant forms of P-glycopro- tein-A52 were solubilized with SDS, treated either with (1) or without (2) endoglycosidase H and then subjected to immunoblot analysis with monoclonal antibody A52 as described under “Experimental Proce- dures.” Bands migrating at 170, 150, and 140 kDa represent mature, core-glycosylated, and unglycosylated forms of P-glycoprotein, respectively.

Article Snippet: The samples were divided into two equal portions, which were then incubated either with (1) or without (2) endoglycosidase Hf (100 units, New England Biolabs, Inc.) for 15 min at room temperature.

Techniques: Expressing, Mutagenesis, Western Blot