<<< Full experiment listing

PXD028965

PXD028965 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleGetting more out of FLAG-Tag co-immunoprecipitation mass spectrometry experiments using FAIMS.
DescriptionCo-immunoprecipitation of proteins coupled to mass spectrometry is critical for the understanding of protein interaction networks. In instances where a suitable antibody is not available, it is common to graft synthetic tags onto target protein sequences and allowing the use of commercially available antibodies for affinity purification. A common approach is through FLAG-Tag co-immunoprecipitation. To allow the selective elution of protein complexes, competitive displacement using a large molar excess of the tag peptides is often carried out. Yet, this creates downstream challenges for the mass spectrometry analysis due to the presence of large quantities of these peptides. Here, we demonstrate that Field Asymmetric Ion Mobility Spectrometry (FAIMS), a gas phase ion separation device prior to mass spectrometry analysis can be applied to FLAG-Tag co-immunoprecipitation experiment to increase the depth of protein coverage. By excluding these abundant tag peptides, we were able to observe deeper coverage of interacting proteins and as a result, deeper biological insights, without the need for additional sample handling or altering sample preparation protocols.
HostingRepositoryPRIDE
AnnounceDate2024-10-08
AnnouncementXMLSubmission_2024-10-08_11:09:23.298.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterChing-Seng Ang
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListacetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
InstrumentOrbitrap Eclipse; Orbitrap Fusion Lumos; Orbitrap Exploris 480
Dataset History
RevisionDatetimeStatusChangeLog Entry
02021-10-06 06:05:23ID requested
12024-10-08 11:09:24announced
Publication List
Ang CS, Sacharz J, Leeming MG, Nie S, Varshney S, Scott NE, Williamson NA, Getting more out of FLAG-Tag co-immunoprecipitation mass spectrometry experiments using FAIMS. J Proteomics, 254():104473(2022) [pubmed]
10.1016/j.jprot.2021.104473;
Keyword List
submitter keyword: FAIMS,Co-IP mass spectrometry, FLAG Tag, Proteomics, ion mobility
Contact List
Nicholas A. Williamson
contact affiliationBio21 Mass Spectrometry and Proteomics Facility, The University of Melbourne, Parkville, Victoria 3052, Australia
contact emailnawill@unimelb.edu.au
lab head
Ching-Seng Ang
contact affiliationUniversity of Melbourne
contact emailcang@unimelb.edu.au
dataset submitter
Full Dataset Link List
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://ftp.pride.ebi.ac.uk/pride/data/archive/2023/02/PXD028965
PRIDE project URI
Repository Record List
[ + ]