PXD034925

PXD034925 is an original dataset announced via ProteomeXchange.

Title	Quantitative proteomics for efficient differentiation of human induced pluripotent stem cells into neural stem cells using data independent acquisition mass spectrometry.
Description	Human induced pluripotent stem cells were differentiated into NSC for 7 days. Cells on days 0, 1, 2, 3, 5 and 7 of differentiation (2.0 × 10⁶) were washed twice with DPBS (−) and then mixed with 100 μL of dissolution buffer consisting of 12 mM sodium deoxycholate, 12 mM N-lauroylsarcosine, 100 mM Tris-HCl pH 9.0, 1% O-GlcNAcase inhibitor, 1% Halt™ protease, and phosphatase inhibitor cocktail. The harvested cells were incubated at 95°C for 5 min and then sonicated to remove the viscosity. One hundred micrograms of protein was diluted to 2.5 μg/μL with dissolution buffer, and the proteins were reduced by adding 10 mM 2-mercaptoetanmol and incubating the mixture at 37℃ for 30 min, followed by another incubation with 20 mM acrylamide at 37℃ in the dark for 60 min. One microgram of mass spectrometry grade lysyl endopeptidase was added to the solution and incubated at 37℃ for 60 min. Then, 2 μg of trypsin was added to the solution and incubated at 37℃ for 6 h. Add an equal amount of ethyl acetate and 0.1% formic acid. After stirring, the supernatant was removed after centrifugation at 14,000 xg. The supernatant was desalted with an Oasis PriME HLB 1 cc Extraction Cartridge according to the manufacturer’s instructions. The protein fraction was dried using a Speed Vac concentrator, and the dried sample was reconstituted with 0.1% FA/3% ACN and analyzed by an Q-Exactive mass spectrometer coupled with a nanoLC instrument.
HostingRepository	jPOST
AnnounceDate	2022-06-27
AnnouncementXML	Submission_2023-08-25_01:33:05.169.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Takaya Urasawa
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	S-carboxamidoethyl-L-cysteine; L-methionine sulfoxide; deamidated L-asparagine
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-06-26 20:30:16	ID requested
1	2022-06-26 20:30:19	announced
2	2022-09-18 03:45:47	announced	2022-09-18: Updated FTP location.
⏵ 3	2023-08-25 01:33:05	announced	2023-08-25: Updated PubMed.

Urasawa T, Koizumi T, Kimura K, Ohta Y, Kawasaki N, Quantitative Proteomics for the Development and Manufacturing of Human-Induced Pluripotent Stem Cell-Derived Neural Stem Cells Using Data-Independent Acquisition Mass Spectrometry. J Proteome Res, 22(6):1843-1854(2023) [pubmed]

submitter keyword: iPS cells, Neural stem cells, Neural differentiation, DIA-MS

Nana Kawasaki
lab head
Takaya Urasawa
contact affiliation	Yokohama city 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/JPST001675/