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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.

SUBMITTER: Schmudlach A

PROVIDER: S-EPMC4764456 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature

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

Schmudlach Andrew A Felton Jeremy J Cipolla Cynthia C Sun Liangliang L Kennedy Robert T RT Dovichi Norman J NJ

The Analyst 20160301 5

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 ...[more]

PMID: 26863548

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