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 35S-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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Guest, P.C. (2017). Two Dimensional Gel Electrophoresis of Insulin Secretory Granule Proteins from Biosynthetically-Labeled Pancreatic Islets. In: Guest, P.C. (eds) Multiplex Biomarker Techniques. Methods in Molecular Biology, vol 1546. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-6730-8_15
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DOI: https://doi.org/10.1007/978-1-4939-6730-8_15
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-6729-2
Online ISBN: 978-1-4939-6730-8
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