PXD075011

PXD075011 is an original dataset announced via ProteomeXchange.

Title	miRNA pull-down and liquid chromatography-mass spectrometry techniques reveal that RBP-binding proteins that interact with miRNAs
Description	To improve protein solubilization, the miRNA pull-down samples were supplemented with sodium dodecyl sulfate (SDS) to a final concentration of 1% (w/v), followed by reduction of disulfide bonds and alkylation of cysteine residues. Proteins were then precipitated by the dropwise addition of 6 volumes of chloroform–acetone (1:2, v/v) under vigorous vortexing, and the mixture was incubated at –20 °C overnight to ensure complete precipitation. The precipitate was collected by centrifugation at 20,000 × g for 30 min at 4 °C, the supernatant was discarded, and the pellet was washed once with ice-cold acetone and air-dried at room temperature. To achieve exhaustive proteolysis, a two-step tryptic digestion strategy was employed. In the first step (aimed at facilitating pellet solubilization and partial digestion), the dried protein pellet was resuspended in 50 mM Tris-HCl buffer (pH 8.5) containing trypsin at an enzyme‑to‑substrate ratio of 1:40 (w/w). The suspension was incubated at 37 °C for 6 h with orbital shaking at 500 rpm in a ThermoMixer. For complete digestion, a second aliquot of trypsin (1:40, w/w) was added, the total volume was adjusted to 50 μL, and incubation was continued overnight (12 h) at 37 °C under constant agitation. The enzymatic reaction was quenched by addition of formic acid to a final concentration of 1% (v/v). The mixture was then centrifuged at 20,000 × g for 30 min at 4 °C, and the resulting supernatant was collected for subsequent LC‑MS/MS analysis.
HostingRepository	iProX
AnnounceDate	2026-02-27
AnnouncementXML	Submission_2026-02-27_18:49:02.748.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Hangao Xie
SpeciesList	scientific name: Sus scrofa domesticus; NCBI TaxID: 9825;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF-X

Revision	Datetime	Status	ChangeLog Entry
0	2026-02-27 18:48:39	ID requested
⏵ 1	2026-02-27 18:49:03	announced

Dataset with its publication pending

submitter keyword: miR-130a

Ruiai Chen
contact affiliation	South China Agricultural University
contact email	chensa727@vip.126.com
lab head
Hangao Xie
contact affiliation	South China Agricultural University
contact email	zpicxhg@163.com
dataset submitter

iProX dataset URI