<<< Full experiment listing

PXD035731

PXD035731 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleiSDAC: in situ digestion of alcohol-fixed cells
DescriptionCurrently, the bottom-up approach, in which proteins are digested by enzymes such as trypsin prior to mass spectrometry, is the mainstream approach in mass spectrometer-based proteomics. In this approach, the enzymatic digestion process strongly affects the reproducibility of protein identification and quantification. Here, we quantitatively evaluated the enzymatic digestion of proteins under various conditions by quantitative proteomics using data-independent acquisition and found that proteins precipitated with acetone after solubilization with SDS were fully digestible without re-solubilization. This result implies that organic solvent treatment makes cells amenable to trypsin digestion. Direct trypsin digestion of methanol-fixed cells achieved the same digestion efficiency and quantitative reproducibility as the conventional method. Furthermore, this method was found to be equally applicable to mouse liver samples. The establishment of this method indicates that the sample preparation process in bottom-up proteomics can be simplified while maintaining high digestion efficiency and is expected to become a general method for sample preparation in bottom-up proteomics in the future.
HostingRepositoryjPOST
AnnounceDate2022-12-16
AnnouncementXMLSubmission_2022-12-16_01:28:24.456.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterAtsushi Hatano
SpeciesList scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListS-carboxamidomethyl-L-cysteine; L-methionine sulfoxide
Instrumentinstrument
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-08-02 05:55:52ID requested
12022-12-16 01:28:24announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: Trypsin, Sample preparation, Methanol-fixation
Contact List
Masaki Matsumoto
lab head
Atsushi Hatano
contact affiliationDepartment of Omics and Systems Biology, Niigata University
dataset submitter
Full Dataset Link List
jPOST 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://ftp.jpostdb.org/JPST001801/