PXD078809-1

PXD078809 is an original dataset announced via ProteomeXchange.

Title	Immunoprecipitation-mass spectrometry (IP-MS) of C5preVII-interacting proteins in 293T cells
Description	HEK-293T cells were seeded in 12-well plates and transfected with C5preVII-HA-expressing plasmid or empty vector. At 24 h after transfection, cells were lysed with ice-cold IP lysis buffer (Invitrogen, Cat#87787) containing protease inhibitor cocktail. The precipitates were removed by centrifugation at 15,000 g at 4 °C for 15 min. For Co-IP analysis, 50 g cell lysate was incubated with 10 L of anti-HA magnetic beads (Thermo, Cat#88836) at room temperature for 1 h with rotation. The beads were washed three times with TBST buffer (25 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.05% Tween-20) and then twice with PBS to remove Tween, followed by resuspension in 2M urea. DTT was added to a final concentration of 2 mM, and the mixture was incubated with shaking at 37 °C for 1 h. Then, iodoacetamide (IAM) was added to a final concentration of 4 mM, and the mixture was incubated at room temperature in the dark for 30-40 min. Trypsin was then added and the sample was digested at 37 °C for 3 h. The digested sample was purified, desalted and analyzed by mass spectrometry. The MS analysis was performed on an EASY-nLC 1200 system coupled to a Q Exactive HF (Thermo Scientific) at the High Throughput Screening Platform at Shanghai Institute for Advanced Immunochemical Studies at ShanghaiTech University. The potential interaction partners of C5preVII were extracted and identified using the Uniprot human protein database (release 2022_04,20613 sequence). The interacting proteins were subsequently confirmed by western blotting combined with Co-IP.
HostingRepository	iProX
AnnounceDate	2026-05-25
AnnouncementXML	Submission_2026-05-25_02:00:13.169.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Peihong Yu
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2026-05-25 01:59:45	ID requested
⏵ 1	2026-05-25 02:00:13	announced

Dataset with its publication pending

submitter keyword: C5preVII, IP-MS, 293T

Jia Liu
contact affiliation	Multi-Omics Core Facility at School of Life Science and Technology, ShanghaiTech University
contact email	liujia@shanghaitech.edu.cn
lab head
Peihong Yu
contact affiliation	Multi-Omics Core Facility at School of Life Science and Technology, ShanghaiTech University
contact email	yuph2022@shanghaitech.edu.cn
dataset submitter

iProX dataset URI