Title: Milieu-induced, selective aggregation of regulated secretory proteins in the trans-Golgi network Document date: 1991_12_2
ID: syyi2ysq_15
Snippet: Membrane Fermeabilizadon . The pellet containing [35S] sulfate-labeled TGN vesicles (-I 141 packed volume) was resuspended in 100141 of buffer containing 0.5 mg/ml saponin and either 10 mM MES-NaOH, pH 7.4, 30 mM KC1, 1 .2 mM leupeptin (nonaggregative milieu) or 10 mM MES-NaOH, pH 6.4, 10 mM CaC12, 1 .2 mM leupeptin (aggregative milieu) . After -15 min incubation at 0°C, samples were subjected to centrifugation (23,000 g for 30 min or 110,000 g .....
Document: Membrane Fermeabilizadon . The pellet containing [35S] sulfate-labeled TGN vesicles (-I 141 packed volume) was resuspended in 100141 of buffer containing 0.5 mg/ml saponin and either 10 mM MES-NaOH, pH 7.4, 30 mM KC1, 1 .2 mM leupeptin (nonaggregative milieu) or 10 mM MES-NaOH, pH 6.4, 10 mM CaC12, 1 .2 mM leupeptin (aggregative milieu) . After -15 min incubation at 0°C, samples were subjected to centrifugation (23,000 g for 30 min or 110,000 g for 15 min), and the entire pellet and supernatant were analyzed by SDS-PAGE and fluorography.
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