PXD036055

PXD036055 is an original dataset announced via ProteomeXchange.

Title	TMT-based quantitative proteomic analysis of the Paphiopedilum barbigerum
Description	Briefly, the frozen samples were ground into a fine powder under liquid nitrogen. Then, the powder was transferred into a centrifuge tube and sonicated on ice three times in lysis buffer consisting of 8 M urea, 2 mM EDTA, 10 mM dithiothreitol (DTT) and 1% protease inhibitor cocktail. The sediment was translated by centrifugation at 12 000×g for 20 min. The protein in the supernatant was deposited with 15% pre-cooled TCA buffer for 2 h at -20℃. The supernatant was abandoned and then residual sediment was rinsed with cold acetone. Finally, the protein was redissolved in a buffer consisting of 8 M urea/100 mM Tetraethylammonium Bromide, pH 8.0). The amount of protein in each sample was measured by the Bradford method and examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protein sample was subjected to a reduction reaction 10 Mm DTT for 1 h at 37℃, and then followed by an alkylation reaction at room temperature for 45 min by incubation in 40 mM iodoacetamide in the darkness (Sohail et al., 2022). Subsequently, the protein solution was diluted with 100 mM TEAB to the urea concentration below 2 M. Trypsin was added at a ratio of 1:50 (enzyme: protein, w/w) for the firstly digestion overnight and then was secondly digested for 4 h at a ratio of 1:100. After digesting, the protein samples were subjected to peptide desalting via a Strata X C18 SPE column and vacuum dried. The peptide was reconstituted with 0.5 M TEAB and conducted in accordance with the manufacturer’s protocol for the TMT kit. In short, one unit of TMT reagent was uncongealed and reconstituted in acetonitrile. The peptides mixtures were incubated at room temperature for 2 h, and merged, desalted and dried by vacuum centrifugation
HostingRepository	iProX
AnnounceDate	2022-08-15
AnnouncementXML	Submission_2022-08-15_05:52:59.318.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Juncai Wang
SpeciesList	scientific name: Paphiopedilum barbigerum; NCBI TaxID: 53071;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-08-15 05:52:17	ID requested
⏵ 1	2022-08-15 05:52:59	announced

Dataset with its publication pending

submitter keyword: Paphiopedilum barbigerum, TMT, proteome profiling

Xiaofeng Liao
contact affiliation	Guizhou Academy of Sciences
contact email	lxfnsd@163.com
lab head
Juncai Wang
contact affiliation	Guizhou Academy of Sciences
contact email	wangjuncai12@126.com
dataset submitter

iProX dataset URI