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Analysis of SCAP N-glycosylation and Trafficking in Human Cells

Article DOI: 10.3791/54709-v • 11:27 min • November 8th, 2016
November 8th, 2016 •
1, 1, 1, 1, 1, 1, 1
1Department of Radiation Oncology, The Ohio State University Comprehensive Cancer Center and College of Medicine

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Summary
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We describe a modified method for membrane fraction isolation from human cells and sample preparation for the detection of SCAP N-glycosylation and total protein by using western blot. We further introduce a GFP-labeling method to monitor SCAP trafficking using confocal microscopy. This protocol can be used in regular biology laboratories.

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SCAP N-glycosylation Trafficking Human Cells Analysis Protocol Conventional Method Lectins PNGase Cancer Cardiovascular Diseases Metabolic Syndromes U87 Cells DMEM FBS Glucose Cell Membrane Fractions Nuclear Extracts PBS
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