PXD035229

PXD035229 is an original dataset announced via ProteomeXchange.

Title	Purification of Biotinylated Peptides using SDB-SCX StageTip
Description	We tried purification of biotinylated peptides using SDB-SCX StageTip to reduce interference ions. We compared this method with the original and optimized workflows. We assessed reproducibility of each enrichment process by three technical replicates. Original workflow with SDB-SCX: A 15-µL slurry of MagCapture HP Tamavidin 2-REV magnetic beads per sample was washed three times with TBS2 (50 mM Tris-HCl, pH 7.5, and 150 mM NaCl). The digested peptides containing 0.1% RapiGest SF were diluted 5-fold with TBS2 and incubated with the Tamavidin 2-REV beads in the presence of 1 mg/mL Pefabloc SC for 3 h at 4 °C. After washing five times with TBS2, biotinylated peptides were eluted with 100 μL of 1 mM biotin in TBS2 for 15 min at 95 °C twice. The combined eluates were subjected to GL-Tip SDB-SCX. After desalting with the first-stage SDB, biotinylated peptides were eluted from the second-stage SCX with 500 mM ammonium acetate and 30% ACN, evaporated in a SpeedVac concentrator, and redissolved in 0.1% TFA and 3% ACN.
HostingRepository	jPOST
AnnounceDate	2022-08-25
AnnouncementXML	Submission_2022-09-18_03:10:06.792.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kohei Nishino
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	S-carboxamidomethyl-L-cysteine; N6-biotinyl-L-lysine; L-methionine sulfoxide; alpha-amino acetylated residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2022-07-11 08:06:40	ID requested
1	2022-08-24 08:00:20	announced
⏵ 2	2022-09-18 03:10:07	announced	2022-09-18: Updated FTP location.

Nishino K, Yoshikawa H, Motani K, Kosako H, Optimized Workflow for Enrichment and Identification of Biotinylated Peptides Using Tamavidin 2-REV for BioID and Cell Surface Proteomics. J Proteome Res, 21(9):2094-2103(2022) [pubmed]

submitter keyword: Biotinylated Peptides, Tamavidin 2-REV, BioID, SDB-SCX

Hidetaka Kosako
lab head
Kohei Nishino
contact affiliation	Tokushima University
dataset submitter

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/JPST001689/