PXD007617-3

PXD007617 is an original dataset announced via ProteomeXchange.

Title	FLAG-IP-UBL3
Description	MDA-MB-231 cells were transfected with plasmids using Lipofectamine 2000. After 24 hr, cells were washed and collected by scraping into ice-cold PBS, pelleted by centrifugation at 400 × g for 1 min at 4°C, and lysed with 1% Triton buffer for 20 min at 4°C. Crude nuclei and unbroken cells were excluded by centrifugation at 20,000 × g for 5 min at 4°C. As a preclean process, the supernatants were incubated twice with 40 μL Protein G-sepharose beads (GE Health care, 50% slurry in lysis buffer) for 1 hr at 4°C. The supernatants were incubated with 50 μl anti-FLAG M2 affinity gel (Sigma, 50% slurry in lysis buffer) for 16hr at 4°C. The beads were washed with 1% Triton buffer and then three times with wash buffer (50mM Tris-HCl [pH7.5]) to remove detergent, and resuspended in 200 μL digestion buffer (2M urea, 50mM Tris-HCl [pH 7.4], 1mM DTT, 5mM iodoacetamide, 1 μg of trypsin [Promega, V5280]), and incubated by shaking at 1,200 rpm for 16hr at 37°C. The samples were acidified with 22 μL of 10% trifluoroacetic acid (TFA). The supernatant containing peptides were then taken by centrifugation at 4,400 × g for 30 sec. The peptides were desalted by C18Tip (Thermo, StageTips), dried by speedvac, and then resuspended in mass spectrometry buffer (2% acetonitrile, 0.2% TFA).
HostingRepository	jPOST
AnnounceDate	2018-09-03
AnnouncementXML	Submission_2021-02-04_17:16:56.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Hiroshi Ageta
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	S-carboxamidomethyl-L-cysteine; L-methionine sulfoxide; alpha-amino acetylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2017-08-31 22:41:06	ID requested
1	2018-09-02 08:00:05	announced
2	2018-09-19 00:28:28	announced	Updated CV, FTP location and Modification list.
⏵ 3	2021-02-04 17:16:57	announced	2021-02-04: Updated FTP location.
4	2022-09-18 03:56:56	announced	2022-09-18: Updated FTP location.

Dataset with its publication pending

submitter keyword: LFQ, MDA-MB-231

Kunihiro Tsuchida
lab head
Hiroshi Ageta
contact affiliation	Fujita Health 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.biosciencedbc.jp/archive/jpostrepos/JPST000315