Chapter title |
Two Dimensional Gel Electrophoresis of Insulin Secretory Granule Proteins from Biosynthetically-Labeled Pancreatic Islets.
|
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Chapter number | 15 |
Book title |
Multiplex Biomarker Techniques
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6730-8_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6729-2, 978-1-4939-6730-8
|
Authors |
Paul C. Guest |
Editors |
Paul C. Guest |
Abstract |
Pulse-chase radiolabeling of cells with radioactive amino acids is a common method for tracking the biosynthesis of proteins. Radiolabeled newly synthesized proteins can be analyzed by a number of techniques such as two dimensional gel electrophoresis (2DE). This chapter presents a protocol for the biosynthetic labeling of pancreatic islets with (35)S-methionine in the presence of basal and stimulatory concentrations of glucose, followed by subcellular fractionation to produce a secretory granule fraction and analysis of the granule protein contents by 2DE. This provides a means of determining whether or not the biosynthetic rates of the entire granule constituents are coordinately regulated. |
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