PXD007617-1

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_2018-09-02_08:00:04.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