MS for BIRC6 Complex from HEK293F
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ABSTRACT: The plasmids were then transiently transfected into HEK293F expression system for overexpression. After incubation for 48 h, the cells were collected, washed one with ice-cold PBS. The resuspended cells were then lysed by sonication, and the lysate supernatant after centrifugation was incubated with M2-FLAG affinity beads for 1.5 h. After several round of washing with buffer B, the FLAG-affinity proteins were eluted by 0.5 mg/mL FLAG peptide dissolved in buffer C. The elute was further purified by gel filtration (Superose 6 Increase Columns, Cytiva) using buffer C. The fraction at 0.5 mL/tube was collected starting from 7 mL. Concentrate a few tubes from the main peak diagram. The purified FLAG-BIRC6 complex was analyzed by mass spectrometry following separation of the complex by SDS-PAGE. The full lane was sliced into 6 pieces from top to bottom with assigned sample numbers: 1, 2-1, 2-2, 3-1, 3-2, and 4.
INSTRUMENT(S): Q Exactive Plus
ORGANISM(S): Homo Sapiens (ncbitaxon:9606)
SUBMITTER: Xiao-Bo Qiu
PROVIDER: MSV000091953 | MassIVE | Mon May 15 17:16:00 BST 2023
SECONDARY ACCESSION(S): PXD042246
REPOSITORIES: MassIVE
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