Preparation of Proliferated Bovine Primary Skeletal Muscle Cells for Bottom-Up Proteomics by LC-MSMS Analysis

Eva Veiseth-Kent; Vibeke Høst; Mona E Pedersen

doi:10.1007/978-1-4939-8897-6_15

Preparation of Proliferated Bovine Primary Skeletal Muscle Cells for Bottom-Up Proteomics by LC-MSMS Analysis

Methods Mol Biol. 2019:1889:255-266. doi: 10.1007/978-1-4939-8897-6_15.

Authors

Eva Veiseth-Kent¹, Vibeke Høst², Mona E Pedersen²

Affiliations

¹ Nofima AS, Osloveien 1, NO-1433, Aas, Norway. eva.veiseth-kent@nofima.no.
² Nofima AS, Osloveien 1, NO-1433, Aas, Norway.

PMID: 30367419
DOI: 10.1007/978-1-4939-8897-6_15

Abstract

This chapter outlines a method for sample preparation for bottom-up proteomics by LC-MSMS analysis of in vitro proliferated bovine primary skeletal muscle cells. The protocol describes the isolation of bovine primary skeletal muscle cells, extraction of proteins, proteolytic digestion of proteins, and desalting of the final peptide samples. The final peptide samples can be analyzed using various LC-MSMS systems after reconstitution in a suitable elution buffer.

Keywords: Isolation of primary muscle cells; LC-MSMS; Peptide generation; Protein extraction; Proteomics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cattle
Cell Separation / methods
Chromatography, Liquid*
Muscle Fibers, Skeletal / cytology
Muscle Fibers, Skeletal / metabolism*
Peptides / metabolism
Proteome*
Proteomics* / methods
Tandem Mass Spectrometry*
Workflow

Substances

Peptides
Proteome