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Chapter title |
Multiplex Sequential Immunoprecipitation of Insulin Secretory Granule Proteins from Radiolabeled Pancreatic Islets.
|
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Chapter number | 14 |
Book title |
Multiplex Biomarker Techniques
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6730-8_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6729-2, 978-1-4939-6730-8
|
Authors |
Paul C. Guest |
Editors |
Paul C. Guest |
Abstract |
Pulse radiolabeling of cells with radioactive amino acids is a common method for tracking the biosynthesis of proteins. Specific proteins can then be immunoprecipitated and analyzed by electrophoresis and imaging techniques. This chapter presents a protocol for the biosynthetic labeling of pancreatic islets with (35)S-methionine, followed by multiplex sequential immunoprecipitation of insulin and three other secretory granule accessory proteins. This provided a means of distinguishing those pancreatic islet proteins with different biosynthetic rates in response to the media glucose concentrations. |