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PXD071203

PXD071203 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleCD4+ T cell surface biotinylation using pre-complexed Siglec-10 Fc and Protein A-APEX2
DescriptionFollicular CD4+ T cells were enriched from pooled splenocytes in immunized C57BL/6J mice. 1 million cells per well in a 96-well V-bottom plate were incubated with 1X PBS containing 2 ug/mL Siglec-10 Fc (or Siglec-10 R119A Fc) pre-complexed with hemin-containing Protein A-APEX2 (1:2 molar ratio) for 30 min at RT, washed with 1X PBS and incubated with 500 uM biotin-tyramide (Millipore) in 1X PBS for 30 min at RT. Cells were then washed with 1X PBS and incubated with 500 uM H2O2 for 1 min at RT to initiate protein biotinylation. Reaction was quenched by washing cells 3 times with 1X PBS supplemented with 5 mM Trolox (Millipore) and 10 mM sodium ascorbate. Cell pellets were lysed in RIPA lysis buffer with protease inhibitors and underwent automated streptavidin beads enrichment (Pierce PI88817) using KingFisher Duo Prime and on-bead trypsin digestion. 10R: Sig-10 R119A Fc; 10W: Sig-10 WT Fc.
HostingRepositoryMassIVE
AnnounceDate2025-11-25
AnnouncementXMLSubmission_2025-11-25_11:31:30.706.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterOlivier Julien
SpeciesList scientific name: Mus musculus; common name: house mouse; NCBI TaxID: 10090;
ModificationListCarbamidomethyl; Oxidation
InstrumentOrbitrap Exploris 480
Dataset History
RevisionDatetimeStatusChangeLog Entry
02025-11-25 11:20:40ID requested
12025-11-25 11:31:31announced
Publication List
no publication
Keyword List
submitter keyword: Siglec-G, B cells, APEX, Cell surface biotinylation, Proximity labeling, DatasetType:Proteomics
Contact List
Olivier Julien
contact affiliationUniversity of Alberta
contact emailojulien@ualberta.ca
lab head
Matthew Macauley
contact affiliationUniversity of Alberta
contact emailmacauley@ualberta.ca
lab head
Olivier Julien
contact affiliationUniversity of Alberta
contact emailojulien@ualberta.ca
dataset submitter
Full Dataset Link List
MassIVE dataset URI
Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://massive-ftp.ucsd.edu/v11/MSV000100018/