PXD062009-1

PXD062009 is an original dataset announced via ProteomeXchange.

Title	APEX2 reaction, biotinylated protein capture and mass spectrometry of TLK2 protein
Description	The APEX2-TLK2 transfected cells were incubated with 500 mM biotin-phenol for 30 minutes in the incubator. Equivoluminal 2 mM media-diluted H2O2 was added to the cells to initiate the labeling reaction for 30 seconds and was removed immediately. The reaction was further quenched by washing three times with ice-cold quenching buffer. Cells were scraped in quenching buffer with protease inhibitors, lysed in RIPA buffer supplemented with 10 mM sodium ascorbate, 1 mM sodium azide, 1 mM Trolox, 1 mM DTT, and protease inhibitors. Samples were briefly sonicated, spun down at 10,000 × g for 10 minutes. The supernatant was incubated with Streptavidin Magnetic Beads at 4°C overnight. The following day, the beads were washed four times with RIPA buffer, two times with 1M KCl solution, two times with 50 mM NH4HCO3 solution, two times with 2 M Urea solution, three times with RIPA buffer, and finally two times with PBS buffer. The beads were then sent to the company (Chi Biomedicine) for digestion and analyzed by Orbitrap Exploris 480 with a FAIMS coupled to an EASY-nanoLC 1200 system under data independent acquisition (DIA) mode with hybrid data strategy.
HostingRepository	iProX
AnnounceDate	2025-03-18
AnnouncementXML	Submission_2025-06-30_18:37:58.322.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Zifeng Wang
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	biotinylated residue
Instrument	Orbitrap Exploris 480; LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2025-03-18 20:20:03	ID requested
⏵ 1	2025-06-30 18:37:58	announced

Dataset with its publication pending

submitter keyword: TLK2, APEX2, DIA

Zifeng Wang
contact affiliation	Sun Yat-sen University
contact email	wzifeng@mail.sysu.edu.cn
lab head
Zifeng Wang
contact affiliation	Sun Yat-sen University
contact email	wzifeng@mail.sysu.edu.cn
dataset submitter

iProX dataset URI