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Proteomics of SEC and IEX fractionated Mouse Embryonic Stem Cells

ABSTRACT: Mouse embryonic stem cells were lysed in Pierce IP Lysis buffer and 2 mg of the clarified extract was fractionated on an HPLC SEC column and 2 mg was fractionated on an HPLC IEX mixed bed column. All fractions were reduced, alkylated, and trypsin digested for mass spectrometry as in Mallam et al. 2019 (PMID: 31665645). Data were collected on an Orbitrap Fusion mass spectrometer using a data dependent 75 minute topspeed method. Dissociation for SEC fractions was by CID, and for IEX fractions was by HCD. RAW files were converted to mzXML and searched using an MSBlender analysis pipeline.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Edward Marcotte

PROVIDER: MSV000091831 | MassIVE | Mon May 01 15:20:00 BST 2023

SECONDARY ACCESSION(S): PXD041915

REPOSITORIES: MassIVE

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