Title: trans-Golgi retention of a plasma membrane protein: mutations in the cytoplasmic domain of the asialoglycoprotein receptor subunit H1 result in trans-Golgi retention Document date: 1995_7_2
ID: tedj3xxz_16
Snippet: The surface/intraeellular distribution of H1 variants was assayed by treating intact cells with 1 mg/ml proteinase K in PBS containing 5 mM EDTA for 30 min at 4°C or 37°C. Digestion was stopped by adding 2 mM PMSF. Digested and control ceils were lysed in gel sample buffer and subjected to SDS-gel electrophoresis and immunoblot analysis using l~I-iodinated protein A. Autoradiographs were quantitated by densitometric scanning. The high mannose p.....
Document: The surface/intraeellular distribution of H1 variants was assayed by treating intact cells with 1 mg/ml proteinase K in PBS containing 5 mM EDTA for 30 min at 4°C or 37°C. Digestion was stopped by adding 2 mM PMSF. Digested and control ceils were lysed in gel sample buffer and subjected to SDS-gel electrophoresis and immunoblot analysis using l~I-iodinated protein A. Autoradiographs were quantitated by densitometric scanning. The high mannose precursor form was used as an internal standard in calculating the fraction of resistant mature form.
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