Issue 5, 2016

Sample preparation protocol for bottom-up proteomic analysis of the secretome of the islets of Langerhans

Abstract

We evaluate a set of protocols for preparation of the secretome from murine islets of Langerhans for bottom-up proteomic analysis. Of the protocols evaluated, a filter-aided sample preparation based approach using sodium dodecyl sulfate as a detergent to solubilize proteins generated the most protein identifications. A total of 362 protein groups (average of 3.7 peptides/protein) were identified from the secretome using the SDS-FASP protocol; a combination of data from three protocols generated 413 protein group identifications. As expected, the identified proteins included insulin 1 and 2, somatostatin, and glucagon, the four main secreted components from islets. STRING network analysis classified the other proteins as being associated with extracellular exosomes, membrane-bounded vesicles, vesicles, and the extracellular region.

Graphical abstract: Sample preparation protocol for bottom-up proteomic analysis of the secretome of the islets of Langerhans

Supplementary files

Article information

Article type
Paper
Submitted
01 Nov 2015
Accepted
01 Feb 2016
First published
02 Feb 2016

Analyst, 2016,141, 1700-1706

Sample preparation protocol for bottom-up proteomic analysis of the secretome of the islets of Langerhans

A. Schmudlach, J. Felton, C. Cipolla, L. Sun, R. T. Kennedy and N. J. Dovichi, Analyst, 2016, 141, 1700 DOI: 10.1039/C5AN02265G

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