PXD028965 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Getting more out of FLAG-Tag co-immunoprecipitation mass spectrometry experiments using FAIMS. |
Description | Co-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. |
HostingRepository | PRIDE |
AnnounceDate | 2024-10-08 |
AnnouncementXML | Submission_2024-10-08_11:09:23.298.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Ching-Seng Ang |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue |
Instrument | Orbitrap Eclipse; Orbitrap Fusion Lumos; Orbitrap Exploris 480 |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2021-10-06 06:05:23 | ID requested | |
⏵ 1 | 2024-10-08 11:09:24 | announced | |
Publication List
Keyword List
submitter keyword: FAIMS,Co-IP mass spectrometry, FLAG Tag, Proteomics, ion mobility |
Contact List
Nicholas A. Williamson |
contact affiliation | Bio21 Mass Spectrometry and Proteomics Facility, The University of Melbourne, Parkville, Victoria 3052, Australia |
contact email | nawill@unimelb.edu.au |
lab head | |
Ching-Seng Ang |
contact affiliation | University of Melbourne |
contact email | cang@unimelb.edu.au |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD028965
- Label: PRIDE project
- Name: Getting more out of FLAG-Tag co-immunoprecipitation mass spectrometry experiments using FAIMS.