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Western blot analysis of cell surface biotinylation of free sulfhydryl groups using maleimide-PEG11-biotin. Oocytes were injected with cRNAs of wild type and triple mutant HA-hPAT1. Lysates of biotinylated oocytes were incubated with neutravidin beads. For elution, the beads were heated to 95 °C in sample buffer. For immunodetection samples were separated on a 10% SDS gel (8 × 10 cm) followed by incubation with the mouse anti-HA tag antibody.
